SCU Weekly

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Revision as of 09:17, 29 September 2013

Animation

June

6.8~6.15

1. transformation of BBa_B0015，BBa_CO261 to psb1AK3 is successful 2.Transformed pSB1T3,pSB1K3, pSB1C3,pSB1A3, BBa_I13504 and BBa_I13504. 4.the bacteria precipitate transformed with pSB1C3 and pSB1A3 were red after centrifugation, the degree of red decreased in this order: pSB1K32 (pinkred) - pSB1A3 (pink) - pSB1A3(tinge) - pSB1K3(yellow). 5. Chloramphenicol was insoluble，and it was tested to be useless using DH5α. 6.prepare A + ,K + ,T + ,C + plates and antibiotic store liquor. 7.Digested and ligated BBa_CO261, pSB1C3,then transformed them 9.Digestion of BBa_I13504, BBa_B0015, and proved that the transformation was successful

6.16~6.22

1.PCR to amplified 8B but failed 2. Chloramphenicol was tested to be effective 3.transformed pSB1T3, K2-8B BBa_CO261, BBa_B0015, BBa_I13504, pSB1K3(pSB1K3) and pSB1A3(pSB1A3) 4.plasmid extraction of F2622 + BBa_CO261 5.Digested F2622 + BBa_CO261 and pSB1T3 (EcoRⅠand PstⅠ) 6.ligated F2622 + BBa_CO261, BBa_B0015 and pSB1T3

6.23~6.29

1.strain conservation of F2622 + BBa_CO261, pSB1C3 2.prepared ddH2O 3.Added 1µl Dpn Ӏ into pSB1T3(pSB1T3) reaction system when digesting

July

6.30~7.6

1.Digested F2622 + BBa_CO261 and pSB1T3(EcoRⅠand PstⅠ) 2.ligated F2622 + BBa_CO261, BBa_I13504, and pSB1T3, but failed to transform

7.7~7.13

1.plated F2622 + BBa_CO261 + BBa_I13504(1.2.3) and F2622 + BBa_CO261 + BBa_B0015(1.2.3) which were ligated last week but failed. 2.cultured BBa_B0015 1 and 2

7.14~7.20

1.no much experiment

7.21~7.27

1. failed to amplify pSB1K3, pSB1A3, pSB1C3 through PCR 2. transformed K2-3G(RBS + Cre), K5-24B, K5-R0040 and 3G, 24B were successful 3.extracted plasmid of K2-8B, F2622 + BBa_CO261 and only K2-8B was successful 4.Liquid cultured 24B, 3G, R0040, 6O, 7F, 12C and F2622 + BBa_CO261 + BBa_I13504, F2622 + BBa_CO261 + BBa_B0015, 4F. 5. Successfully ligated F2622 + BBa_CO261 + BBa_I13504, F2622 + BBa_CO261 + BBa_B0015

RePCR by taq enzyme, we got experiences: Taq is suitable for amplifying snippets of 2kb in size, and the reaction is fast 10X taq buffer with (NH4)2SO4 reduces nonspecification, B0034M Mg2 + is suitable for taq Avoid DNA pollution during template preparation No pollution of raw material, it would be better to operate on the laminar Negative control is necessary.

7.28~8.2

1.Recovered PCR product of 27th 2.ligated F2622 + BBa_CO261 + BBa_I13504, F2622 + BBa_CO261 + BBa_B0015 with pSB1C3 carrier, then did transformation and liquid culture 3.Liquid cultured R0040, 8B, 6O, F2622 + BBa_CO261, BBa_B0015 4.Replated F2622 + BBa_CO261 + BBa_I13504, F2622 + BBa_CO261 + BBa_B0015 5.Digested R0040, 6O, 24B 6.Ligated 6O with 24B, then did transformation 7.transformed 18O, 19C

August

8.3~8.10

1.ligated 6N + 19C + BBa_B0015, after failing to ligate 6N(T7, promoter) with 19C(reporter) 2.Liquid cultured 6O + 24B, and gel analysis of plasmid extraction and digestion products proved that the ligation was successful 3.Transformed K3- I746361, K3-K145215, K2-15J and 5J + 2C. K145215 and 15J turned out to be a failure. Liquid cultured the rest. 4.Digested 6O, 24B, pSB1C3, and ligated them overnight

8.11~8.17

1.transformed 5J + 2C，18O, 19C， I746361,K145215,15J，K5-J23032，F2622 + BBa_CO261 2.Extracted plasmid of K145215,15J,5J + 2C,F2622 + BBa_CO261 3.Digested K145215，6N + I13507，BBa_CO261，5J + 2C,F2622 + BBa_CO261， 4.Liquid cultured F2622，6N + 19C + BBa_B0015，J23032 5.Sent K145215, 6N + I13507, F2622, 6N + 19C(PSN1C3), 6N + 19C + BBa_B0015(PSB1C3) to biotech company to sequence them.

8.18~8.24

1.Ligated R0040 + J23032(Ptet + lock) ,B0034 + 2C , PO + LOX + R0040 2.Transformed B0034 + 6N,6O + 24B,F2622 + BBa_CO261, R0040 + J23032 3.Liquid cultured 5J + 2C, R0040 + J23032,6O + 24B, 4.Extracted plasmid of 5J + 2C，J23032, B0034（strong RBS）,14G + BBa_B0015, R0040 + J23032, 6O + 24B, 6N + B0034 and then did digestion. 5.Digested cre, po，6O + 24B, B0034,2C. 6.Sent 6O + 24B, 5J + 2C to sequence, however all turned out to be wrong, so we need to religate them. 7.Sent 5J + 2C + PSB1C3，14G + BBa_B0015 + PSB1C3, R0040 + J23032, 6N + B0034 to sequence 8.Amplified I746361 through PCR

8.25~8.31

1. Digested R0040 + J23032 and F2622 + BBa_B0015, B0034 + 2C, 6O + 24B 2.Designed primer of APO 3.Failed to PCR Cre + ssrA 4.Transformed B0034 + 2C, R0040 + PO, 5.Recultured 6O + 24B, F2622 + BBa_CO261 6.Single digested F2622 + BBa_B0015,R0040 + po, compared with previous single digested BBa_B0015, but failed. 7. Redigested BBa_B0015, then did ligation and transformation. 8.Sent to sequence: 14G + BBa_B0015, R0040 + J23032, 6O + 24B, R0040 + po, 6N + B0034, B0034 + 2C 9.Recovered BBa_B0015 and R0040 from conservation

September

9.1~9.7

1. succeeded in amplifying Cre + ssrA by PCR 2.Digested R0040, 6N + B0034, pSB1C3, pSB1K3, pSB1A3, pSB1T3, 6O + 24B, F2622 + BBa_CO261, Cre + BBa_B0015, 6N→BBa_B0015, B0034 + 14G + BBa_B0015, 4F + 6N, PCRed F2622 + BBa_B0015, F2622 + BBa_B0015 + R0040 + J23032, 6G, 24B 3.ligated Cre + BBa_B0015, F2622 + BBa_CO261(pSB1C3), B0034 + 14G + BBa_B0015(pSB1T3), 6N + B0034 + 14G + BBa_B0015, R0040 + J23032 + F2622 + BBa_B0015(pSB1T3), R0040 + po + I13507 (pSB1T3), RBS + Cre + ssrA + B0034 + 14G + BBa_B0015(pSB1T3) (C23 for short), RBS + Cre + ssrA (pSB1C3) (CreS for short), F2622 + BBa_B0015 (pSB1C3) (1223 for short), B0034 + 14G + BBa_B0015(pSB1C3), R0040 + BBa_I13504 (pSB1C3), I746361 + BBa_I13504 (pSB1C3), R0040 → BBa_B0015 (pSB1C3), 6G + 24B (pSB1C3), then transformed them. 4. Cultured 6O + 24B, B0034 + 2C 5.Digested pSB1C3, pSB1K3, pSB1A3, pSB1T3, 6O + 24B, F2622 + BBa_CO261 6.Transformed Cre + BBa_B0015, 18F + 4F, 4F + 6N, 6N + B0034 + 14G + BBa_B0015, pSB1A2, K56G, Plac, C23, 2012K1220, K33L, K3J04650, J04650 7.Transformed pSB1A2, K56G, Plac, C23, 2012K1220, K33L, K3J04650, J04650 8.Sent to sequence: B0034 + 14G + BBa_B0015, 6N → BBa_B0015, B0034 + 2C, 6O + 24B, 12C + BBa_B0015 → 7F, F2622 + BBa_B0015 → 1B0034 9.Did gel extraction of 6N + 19C + BBa_B0015, RBS + Cre + ssrA

9.8~9.14

1.Digested CreS, 1223, B0034 + 14G + BBa_B0015, 6N→BBa_B0015, I13507, pSB1C3, CreD, CreE, R0040 + po, R0040 + po + I13507, 6G + 3L 2.Liquid cultured R0040→BBa_B0015, 6G + 24B 3. Ligated 6G + 3L, R0040 + po + I13507, R0040 + J23032 + J04650 ,R0040 + po + I13507, CreS, C23 4.Sent to sequence: 1223, B0034→BBa_B0015, 6N→BBa_B0015, R0040→BBa_B0015, 6G + 3L, R0040 + po + I13507 5.Traditional assembly of Cre and C23 6.96-well plates experiment of R0040 + po 7.Cultured R0040 + BBa_I13504 and observed its fluorescence 8.Transformed R0040 + J23032 + J04650, 6G + 3L, R0040 + po + I13507

9.15~9.21

1.Miniprep：I746361 + I13507, 6G + 3L, R0040 + J23032 + J04650, R0040 + Po + I13507, I746361 + I13507, Cres + B0034 + 14G + BBa_B0015, I746361 + I13507 + R0040→BBa_B0015, K145215 + R0040 + J23032 + J04650, I746361 + I13507 + R0040 + J23032 + F2622 + BBa_B0015 + K145215, R0040 + po + I13507, 6g, I746361→BBa_B0015 + K145215, C23, I13521, K145215 + R0040 + J23032 + J04650 2.Digestion: I746361 + I13507, R0040 + J23032 + 12K, Cres, 5I + J23032 + J04650, Cre + B0034 + 14G + BBa_B0015, I746361 + I13507 + R0040→BBa_B0015,K145215 + R0040 + J23032 + J04650, R0040 + Po + I13507, R0040 + po + I13507, 6g, I746361→BBa_B0015 + K145215 3.Liquid culture: I746361 + I13507, Cres, I746361 + I13507 + R0040→BBa_B0015, Cre + B0034 + 14G + BBa_B0015, cres + Po + I13507, 6G, R0040 + J23032 + J04650, K145215 + R0040 + J23032 + J04650, R0040, K145215 + B0034 + J23032 + BBa_B0015, R0040 + Po + I13507 4. Prepared A + and T + plates 5. Ligated I746361→BBa_B0015 + K145215 and R0040 + I13507,then did transformation 6. Function test: K145215 + R0040 + J23032 + J04650, R0040 + J23032 + J04650, R0040 + J23032 + J04650 + K145215, I13521, I746361 + I13507, I746361 + I13507 + R0040→BBa_B0015, I746361 + 220 + R0040→BBa_B0015 + K145215, R0040 + Po + I13507 7. Gel extraction of R0040 + po + I13507 and 6g 8. Sent 6G + 3L, R0040 + J23032 + J04650 + K145215, I746361 + I13507 + R0040→BBa_B0015, I746361 + 220 + R0040→BBa_B0015 + K145215 to sequence

9.22-9.28

1.Resent I746361 + 220 + R0040→BBa_B0015 + K145215 to sequence 2.Prepare LB liquid culture and LB plates 3.Liquid cultured 6G + R0040 + PO + I13507, C23,B0034→BBa_B0015 and 3L, 6G→I13507, and then extracted plasmid 4.Digested I13521、C23,B0034 + 14G + BBa_B0015、3L, cre、R0040 + po + I13507 + 6G、6G→I13507、B0034→BBa_B0015、B0034 + 14G + BBa_B0015 + psb1c3 5.Gel extraction of B0034 + 14G + BBa_B0015、3L、B0034 + 14G + BBa_B0015 + psb1c3、cre、3L、6G→I13507 and I13507 6.Function test of I13521、 I746361→BBa_B0015、 I746361→K145215、 I746361 + I13507 7.PCRI13507 8.Ligated cre and B0034→BBa_B0015 9.Transformed c23、B0034→BBa_B0015 and 3L

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 ----
-<html>
+==June==
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+===6.8~6.15===
-@font-face{
+. transformation of BBa_B0015，BBa_CO261 to psb1AK3 is successful
-font-family:"Times New Roman";
+.Transformed pSB1T3,pSB1K3, pSB1C3,pSB1A3, BBa_I13504 and BBa_I13504.
-}
+.the bacteria precipitate transformed with pSB1C3 and pSB1A3  were red after centrifugation, the degree of red decreased in this order: pSB1K32 (pinkred) - pSB1A3 (pink) - pSB1A3(tinge) - pSB1K3(yellow).
+. Chloramphenicol was insoluble，and it was tested to be useless using DH5α.
+.prepare A + ,K + ,T + ,C +  plates and antibiotic store liquor.
+.Digested and ligated  BBa_CO261, pSB1C3,then transformed them
+.Digestion of  BBa_I13504, BBa_B0015, and proved that the transformation was successful
-@font-face{
+===6.16~6.22===
-font-family:"&#23435;&#20307;";
+.PCR to amplified 8B but failed
-}
+. Chloramphenicol was tested to be effective
+.transformed pSB1T3, K2-8B BBa_CO261, BBa_B0015, BBa_I13504, pSB1K3(pSB1K3) and pSB1A3(pSB1A3)
+.plasmid extraction of  F2622 + BBa_CO261
+.Digested F2622 + BBa_CO261 and pSB1T3 (EcoRⅠand PstⅠ)
+.ligated F2622 + BBa_CO261, BBa_B0015 and pSB1T3
-@font-face{
+===6.23~6.29===
-font-family:"&#24494;&#36719;&#38597;&#40657;";
+.strain conservation of  F2622 + BBa_CO261, pSB1C3
-}
+.prepared ddH2O
+.Added 1µl Dpn Ӏ into pSB1T3(pSB1T3) reaction system when digesting
-@font-face{
+==July==
-font-family:"Tahoma";
+===6.30~7.6===
-}
+.Digested F2622 + BBa_CO261 and pSB1T3(EcoRⅠand PstⅠ)
+.ligated F2622 + BBa_CO261, BBa_I13504, and pSB1T3, but failed to transform
-@font-face{
+===7.7~7.13===
-font-family:"Symbol";
+.plated F2622 + BBa_CO261 + BBa_I13504(1.2.3) and F2622 + BBa_CO261 + BBa_B0015(1.2.3) which were ligated  last week but failed.
-}
+.cultured BBa_B0015 1 and 2
-@font-face{
+===7.14~7.20===
-font-family:"Arial";
+.no much experiment
-}
-@font-face{
+===7.21~7.27===
-font-family:"&#40657;&#20307;";
+. failed to amplify pSB1K3, pSB1A3, pSB1C3 through  PCR
-}
+. transformed K2-3G(RBS + Cre), K5-24B, K5-R0040 and 3G, 24B were successful
+.extracted plasmid of K2-8B, F2622 + BBa_CO261 and only K2-8B was successful
+.Liquid cultured 24B, 3G, R0040, 6O, 7F, 12C and F2622 + BBa_CO261 + BBa_I13504, F2622 + BBa_CO261 + BBa_B0015, 4F.
+. Successfully ligated F2622 + BBa_CO261 + BBa_I13504, F2622 + BBa_CO261 + BBa_B0015
-@font-face{
+RePCR by taq enzyme, we got experiences:
-font-family:"Courier New";
+Taq is suitable for amplifying snippets of 2kb in size, and the reaction is fast
-}
+X taq buffer with (NH4)2SO4 reduces nonspecification,
+B0034M Mg2 +  is suitable for taq
+Avoid DNA pollution during template preparation
+No pollution of raw material, it would be better to operate on the laminar
+Negative control is necessary.
-@font-face{
+===7.28~8.2===
-font-family:"Wingdings";
+.Recovered  PCR product of 27th
-}
+.ligated F2622 + BBa_CO261 + BBa_I13504, F2622 + BBa_CO261 + BBa_B0015 with pSB1C3 carrier, then did  transformation and liquid culture
+.Liquid cultured R0040, 8B, 6O, F2622 + BBa_CO261, BBa_B0015
+.Replated F2622 + BBa_CO261 + BBa_I13504, F2622 + BBa_CO261 + BBa_B0015
+.Digested R0040, 6O, 24B
+.Ligated 6O with 24B, then did transformation
+.transformed 18O, 19C
-@font-face{
+== August==
-font-family:"Calibri";
+===8.3~8.10===
-}
+.ligated 6N + 19C + BBa_B0015, after failing to ligate 6N(T7, promoter) with 19C(reporter)
+.Liquid cultured 6O + 24B, and gel analysis of plasmid extraction and digestion products proved that the ligation was successful
+.Transformed K3- I746361, K3-K145215, K2-15J and 5J + 2C. K145215 and 15J turned out to be a failure. Liquid cultured the rest.
+.Digested 6O, 24B, pSB1C3, and ligated them overnight
-@font-face{
+===8.11~8.17===
-font-family:"Cambria Math";
+.transformed 5J + 2C，18O, 19C， I746361,K145215,15J，K5-J23032，F2622 + BBa_CO261
-}
+.Extracted plasmid of K145215,15J,5J + 2C,F2622 + BBa_CO261
+.Digested K145215，6N + I13507，BBa_CO261，5J + 2C,F2622 + BBa_CO261，
+.Liquid cultured F2622，6N + 19C + BBa_B0015，J23032
+.Sent K145215, 6N + I13507, F2622, 6N + 19C(PSN1C3), 6N + 19C + BBa_B0015(PSB1C3) to biotech company to sequence them.
-p.p0{
+===8.18~8.24===
-margin:0pt;
+.Ligated R0040 + J23032(Ptet + lock) ,B0034 + 2C , PO + LOX + R0040
-margin-bottom:0.0001pt;
+.Transformed B0034 + 6N,6O + 24B,F2622 + BBa_CO261, R0040 + J23032
-margin-bottom:0pt;
+.Liquid cultured 5J + 2C, R0040 + J23032,6O + 24B,
-margin-top:0pt;
+.Extracted plasmid of 5J + 2C，J23032, B0034（strong RBS）,14G + BBa_B0015, R0040 + J23032, 6O + 24B, 6N + B0034 and then did digestion.
-text-align:justify;
+.Digested cre, po，6O + 24B, B0034,2C.
-font-size:10.5000pt; font-family:'Calibri'; }
+.Sent 6O + 24B, 5J + 2C to sequence, however all turned out to be wrong, so we need to religate them.
+.Sent 5J + 2C + PSB1C3，14G + BBa_B0015 + PSB1C3, R0040 + J23032, 6N + B0034 to sequence
+.Amplified  I746361 through PCR
-span.10{
+===8.25~8.31===
-font-size:10.0000pt; font-family:'Times New Roman'; }
+. Digested R0040 + J23032 and F2622 + BBa_B0015, B0034 + 2C, 6O + 24B
+.Designed primer of APO
+.Failed to PCR Cre + ssrA
+.Transformed B0034 + 2C, R0040 + PO,
+.Recultured 6O + 24B, F2622 + BBa_CO261
+.Single digested F2622 + BBa_B0015,R0040 + po, compared with previous single digested BBa_B0015, but failed.
+. Redigested  BBa_B0015, then did ligation and transformation.
+.Sent to sequence:  14G + BBa_B0015, R0040 + J23032, 6O + 24B, R0040 + po, 6N + B0034, B0034 + 2C
+.Recovered BBa_B0015 and  R0040 from conservation
-span.15{
+==September==
-font-size:10.0000pt; font-family:'Times New Roman'; }
+===9.1~9.7===
+. succeeded in amplifying Cre + ssrA by PCR
+.Digested R0040, 6N + B0034, pSB1C3, pSB1K3, pSB1A3, pSB1T3, 6O + 24B, F2622 + BBa_CO261, Cre + BBa_B0015, 6N→BBa_B0015, B0034 + 14G + BBa_B0015, 4F + 6N, PCRed F2622 + BBa_B0015, F2622 + BBa_B0015 + R0040 + J23032,  6G, 24B
+.ligated Cre + BBa_B0015, F2622 + BBa_CO261(pSB1C3), B0034 + 14G + BBa_B0015(pSB1T3), 6N + B0034 + 14G + BBa_B0015, R0040 + J23032 + F2622 + BBa_B0015(pSB1T3), R0040 + po + I13507 (pSB1T3), RBS + Cre + ssrA + B0034 + 14G + BBa_B0015(pSB1T3) (C23 for short),  RBS + Cre + ssrA (pSB1C3) (CreS for short), F2622 + BBa_B0015 (pSB1C3) (1223 for short), B0034 + 14G + BBa_B0015(pSB1C3), R0040 + BBa_I13504 (pSB1C3), I746361 + BBa_I13504 (pSB1C3),  R0040 → BBa_B0015 (pSB1C3), 6G + 24B (pSB1C3),  then transformed them.
+. Cultured 6O + 24B, B0034 + 2C
+.Digested pSB1C3, pSB1K3, pSB1A3, pSB1T3, 6O + 24B, F2622 + BBa_CO261
+.Transformed Cre + BBa_B0015, 18F + 4F, 4F + 6N, 6N + B0034 + 14G + BBa_B0015, pSB1A2, K56G, Plac, C23, 2012K1220, K33L, K3J04650, J04650
+.Transformed pSB1A2, K56G, Plac, C23, 2012K1220, K33L, K3J04650, J04650
+.Sent to sequence: B0034 + 14G + BBa_B0015, 6N → BBa_B0015, B0034 + 2C, 6O + 24B,  12C + BBa_B0015 → 7F, F2622 + BBa_B0015 → 1B0034
+.Did gel extraction of 6N  + 19C + BBa_B0015, RBS + Cre + ssrA
-span.16{
+===9.8~9.14===
-font-size:10.0000pt; font-family:'Times New Roman'; }
+.Digested CreS, 1223, B0034 + 14G + BBa_B0015, 6N→BBa_B0015, I13507, pSB1C3, CreD, CreE, R0040 + po, R0040 + po + I13507, 6G + 3L
+.Liquid cultured R0040→BBa_B0015, 6G + 24B
+. Ligated 6G + 3L, R0040 + po + I13507, R0040 + J23032 + J04650 ,R0040 + po + I13507, CreS, C23
+.Sent to sequence: 1223, B0034→BBa_B0015, 6N→BBa_B0015, R0040→BBa_B0015, 6G + 3L, R0040 + po + I13507
+.Traditional assembly of Cre and C23
+.96-well plates experiment of R0040 + po
+.Cultured R0040 + BBa_I13504 and observed its fluorescence
+.Transformed R0040 + J23032 + J04650, 6G + 3L, R0040 + po + I13507
-span.17{
+===9.15~9.21===
-color:rgb(53,161,212); text-decoration:underline ;font-size:10.0000pt; font-family:'Times New Roman'; }
+.Miniprep：I746361 + I13507, 6G + 3L, R0040 + J23032 + J04650, R0040 + Po + I13507,  I746361 + I13507, Cres + B0034 + 14G + BBa_B0015,  I746361 + I13507 + R0040→BBa_B0015, K145215 + R0040 + J23032 + J04650, I746361 + I13507 + R0040 + J23032 + F2622 + BBa_B0015 + K145215, R0040 + po + I13507, 6g,  I746361→BBa_B0015 + K145215, C23, I13521, K145215 + R0040 + J23032 + J04650
+.Digestion: I746361 + I13507, R0040 + J23032 + 12K, Cres, 5I + J23032 + J04650, Cre + B0034 + 14G + BBa_B0015,  I746361 + I13507 + R0040→BBa_B0015,K145215 + R0040 + J23032 + J04650,  R0040 + Po + I13507, R0040 + po + I13507, 6g,  I746361→BBa_B0015 + K145215
+.Liquid culture:  I746361 + I13507, Cres,  I746361 + I13507 + R0040→BBa_B0015, Cre + B0034 + 14G + BBa_B0015, cres + Po + I13507, 6G,  R0040 + J23032 + J04650, K145215 + R0040 + J23032 + J04650, R0040, K145215 + B0034 + J23032 + BBa_B0015, R0040 + Po + I13507
+. Prepared A +     and T +   plates
+. Ligated  I746361→BBa_B0015 + K145215 and R0040 + I13507,then did transformation
+. Function test: K145215 + R0040 + J23032 + J04650, R0040 + J23032 + J04650, R0040 + J23032 + J04650 + K145215, I13521,  I746361 + I13507,  I746361 + I13507 + R0040→BBa_B0015,  I746361 + 220 + R0040→BBa_B0015 + K145215, R0040 + Po + I13507
+. Gel extraction  of R0040 + po + I13507 and 6g
+. Sent 6G + 3L, R0040 + J23032 + J04650 + K145215,  I746361 + I13507 + R0040→BBa_B0015,  I746361 + 220 + R0040→BBa_B0015 + K145215 to sequence
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+===9.22-9.28===
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+.Resent  I746361 + 220 + R0040→BBa_B0015 + K145215 to sequence
+.Prepare LB liquid culture and LB plates
+.Liquid cultured 6G + R0040 + PO + I13507, C23,B0034→BBa_B0015 and 3L, 6G→I13507, and then extracted plasmid
+.Digested I13521、C23,B0034 + 14G + BBa_B0015、3L, cre、R0040 + po + I13507 + 6G、6G→I13507、B0034→BBa_B0015、B0034 + 14G + BBa_B0015 + psb1c3
+.Gel extraction of B0034 + 14G + BBa_B0015、3L、B0034 + 14G + BBa_B0015 + psb1c3、cre、3L、6G→I13507 and I13507
+.Function test of I13521、 I746361→BBa_B0015、 I746361→K145215、 I746361 + I13507
+.PCRI13507
+.Ligated cre and B0034→BBa_B0015
+.Transformed c23、B0034→BBa_B0015 and 3L
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-div.Section0{page:Section0;}</style></head><body style="tab-interval:36pt; " ><!--StartFragment--><div class="Section0"  style="layout-grid:18.0000pt;
-" ><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span><a href="https://2013.igem.org/Team:SCU_China/Notebook" ><span class="17"  style="mso-spacerun:'yes'; color:rgb(53,161,212); text-decoration:underline ;font-size:10.0000pt; font-family:'&#23435;&#20307;'; " >Back&nbsp;to&nbsp;the&nbsp;Protocol&nbsp;Page</span></a></span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.8~6.15</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >Transformation</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;of&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015,&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;to&nbsp;psb1</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >AK</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3&nbsp;is&nbsp;successful</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Transfo</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >r</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >med&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1T3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;pSB1K3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1C3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;pSB1A3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_I13504</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;and&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_I13504</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;T</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >he&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >bacteria&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >precipitate&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >transformed&nbsp;with&nbsp;pSB1C3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;and&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1A3</span><span style="mso-spacerun:'yes'; color:rgb(67,67,67); font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >were&nbsp;red&nbsp;after&nbsp;</span><span><a href="app:ds:centrifugation" ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >centrifugation</span></a></span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;the&nbsp;degree&nbsp;of&nbsp;red&nbsp;decreased&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >in&nbsp;this&nbsp;order:&nbsp;pSB1K3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;(</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >red</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >)&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >-</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;pSB1A3&nbsp;(</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >pink</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >)&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >-</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;pSB1A3(</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >tinge</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >)&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >-</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;pSB1K3(</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >yellow</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >).</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >5.&nbsp;Chloramphenicol&nbsp;was&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >in</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >soluble<font face="&#23435;&#20307;" >&#65292;</font><font face="Times New Roman" >and&nbsp;it&nbsp;was&nbsp;tested&nbsp;to&nbsp;be&nbsp;useless&nbsp;</font></span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >using&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >DH5</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Arial'; " >&#945;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >repare&nbsp;A</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; vertical-align:super; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; vertical-align:super; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,T</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; vertical-align:super; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,C</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; vertical-align:super; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;plates&nbsp;and&nbsp;antibiotic&nbsp;store&nbsp;liquor</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >7.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Digested&nbsp;and&nbsp;assembled&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1C3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >then&nbsp;transformed&nbsp;them</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >9.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Digestion&nbsp;of&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_I13504&nbsp;and</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015,&nbsp;and&nbsp;results&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >proved</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;that&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >the&nbsp;transformation&nbsp;was&nbsp;successful&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.16~6.22</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >PCR&nbsp;to&nbsp;amplif</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >y</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;8B&nbsp;but&nbsp;failed</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.&nbsp;Chloramphenicol&nbsp;was&nbsp;tested&nbsp;to&nbsp;be&nbsp;effective</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;Transformed</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1T3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_I13504</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1K3&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(pSB1K3)&nbsp;and&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1A3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(pSB1A3)</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >lasmid&nbsp;extraction&nbsp;of&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >5.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Digested&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;and&nbsp;pSB1T3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >EcoR&#8544;and&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Pst&#8544;)</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;A</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ssembled&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;and&nbsp;pSB1T3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.23~6.29</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;S</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >train&nbsp;conservation&nbsp;of&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1C3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >repared&nbsp;ddH</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; vertical-align:sub; " >2</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >O&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Added&nbsp;1&#181;l&nbsp;</span><span style="mso-spacerun:'yes'; font-style:italic; font-size:10.5000pt; font-family:'Times New Roman'; " >Dpn</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&#1216;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >into&nbsp;pSB1T3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1T3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >)&nbsp;reaction&nbsp;system&nbsp;when&nbsp;digesting</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.30~7.6</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Digested&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;and&nbsp;pSB1T3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >EcoR&#8544;and&nbsp;Pst&#8544;)&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;A</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ssembled&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_I13504</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;and&nbsp;pSB1T3,&nbsp;but&nbsp;failed&nbsp;to&nbsp;transform</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;them</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >7.7~7.13</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >lated&nbsp;BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261&nbsp;+&nbsp;BBa_I13504&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(1.2.3)&nbsp;and&nbsp;BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261&nbsp;+&nbsp;BBa_B0015&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(1.2.3)&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >which</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;were&nbsp;assembled&nbsp;last&nbsp;week</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;but&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >failed</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;C</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ultured&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;1&nbsp;and&nbsp;2&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >7.14~7.20</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;N</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >o&nbsp;much&nbsp;experiment</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >7.21~7.27</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >F</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ailed&nbsp;to&nbsp;amplify&nbsp;pSB1K3,&nbsp;pSB1A3,&nbsp;pSB1C3&nbsp;through&nbsp;PCR</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >T</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ransfo</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >r</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >med&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_I61046&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(RBS</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Cre),&nbsp;K5-R0040&nbsp;and&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >they&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >were&nbsp;successful</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;E</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >xtracted&nbsp;plasmid&nbsp;of&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1T3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;and&nbsp;only&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1T3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >was&nbsp;successful&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Incubated&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261&nbsp;+&nbsp;BBa_I13504</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >5.&nbsp;Successfully&nbsp;assembled&nbsp;BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261&nbsp;+&nbsp;BBa_I13504</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; color:rgb(255,0,0); font-size:10.5000pt; font-family:'Times New Roman'; " >RePCR&nbsp;by&nbsp;taq&nbsp;enzyme,&nbsp;we&nbsp;got&nbsp;experience</span><span style="mso-spacerun:'yes'; color:rgb(255,0,0); font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >s</span><span style="mso-spacerun:'yes'; color:rgb(255,0,0); font-size:10.5000pt; font-family:'Times New Roman'; " >:</span><span style="mso-spacerun:'yes'; color:rgb(255,0,0); font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p23  style="margin-left:18.0000pt; text-indent:-18.0000pt; margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Taq&nbsp;is&nbsp;suitable&nbsp;for&nbsp;amplif</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >y</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ing&nbsp;snippets&nbsp;of&nbsp;2kb&nbsp;in&nbsp;size,&nbsp;and&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >the&nbsp;reaction&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >is&nbsp;fast</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p23  style="margin-left:18.0000pt; text-indent:-18.0000pt; margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >10X&nbsp;taq&nbsp;buffer&nbsp;with&nbsp;(NH</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; vertical-align:sub; " >4</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >)</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; vertical-align:sub; " >2</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >SO</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; vertical-align:sub; " >4</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;reduces&nbsp;nonspecific</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >tion</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p23  style="margin-left:18.0000pt; text-indent:-18.0000pt; margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >2mM&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Mg2</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;is&nbsp;suitable&nbsp;for&nbsp;taq</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p23  style="margin-left:18.0000pt; text-indent:-18.0000pt; margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >A</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >void&nbsp;DNA&nbsp;pollution&nbsp;during&nbsp;template&nbsp;preparation</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p23  style="margin-left:18.0000pt; text-indent:-18.0000pt; margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >5.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >No&nbsp;pollution&nbsp;of&nbsp;raw&nbsp;material,&nbsp;it&nbsp;would&nbsp;be&nbsp;better&nbsp;to&nbsp;operate&nbsp;on&nbsp;the&nbsp;laminar</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p23  style="margin-left:18.0000pt; text-indent:-18.0000pt; margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >N</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >e</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >gative&nbsp;control&nbsp;is&nbsp;necessary</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p23  style="margin-left:18.0000pt; margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >7.28~8.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >2</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Recovered&nbsp;&nbsp;PCR&nbsp;product&nbsp;of&nbsp;27</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; vertical-align:sub; " >th</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; vertical-align:sub; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;A</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ssembled&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261&nbsp;+&nbsp;BBa_I13504</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;with&nbsp;pSB1C3&nbsp;carrier,&nbsp;then&nbsp;did&nbsp;transfo</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >r</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >mation&nbsp;and&nbsp;incubate</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Incubated&nbsp;R0040,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1T3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >R0062</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Replated&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261&nbsp;+&nbsp;BBa_I13504</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >5.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Digested&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >R0062</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >J61064</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;Assembled&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >R0062with</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >J61064</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;then&nbsp;did&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >transformation</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >8.3~8.10</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >1</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Incubated&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >R0062+</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >J61064</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;and</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;gel&nbsp;analysis&nbsp;of</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;plasmid&nbsp;extraction&nbsp;and&nbsp;digestion</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;products</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;proved&nbsp;that&nbsp;the&nbsp;ligation&nbsp;was&nbsp;successful</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >2</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;Transformed</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Digested&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >R0062</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >J61064</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;pSB1C3,&nbsp;and&nbsp;assembled&nbsp;them&nbsp;overnight</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >8.11~8.17</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;T</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ransformed</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I746361,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_K145215<font face="&#23435;&#20307;" >&#65292;</font></span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032<font face="&#23435;&#20307;" >&#65292;</font><font face="Times New Roman" >BB</font></span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Extracted&nbsp;plasmid&nbsp;of</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Digested&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >,&nbsp;BBa_CO261,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&#65292;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Incubated&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622<font face="&#23435;&#20307;" >&#65292;</font></span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >5.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Sent&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >to</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;biotech&nbsp;company&nbsp;to</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;sequenc</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >e&nbsp;them.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >8.18~8.24</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Assembled&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(Ptet</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >lock),&nbsp;PO</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >LOX</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Transformed&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_R0062&nbsp;+&nbsp;BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Incubated&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_R0062&nbsp;+&nbsp;BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Extracted&nbsp;plasmid&nbsp;of</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032,</span><span style="mso-spacerun:'yes'; color:rgb(255,0,0); font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >B0034</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&#65288;<font face="Times New Roman" >strong&nbsp;RBS</font></span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >,&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_R0062&nbsp;+&nbsp;Cre&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >and&nbsp;then&nbsp;did&nbsp;digestion</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >5.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Digested&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_R0062&nbsp;+&nbsp;BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0034.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Sent&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_R0062&nbsp;+&nbsp;BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >to&nbsp;sequenc</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >e</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;however&nbsp;all&nbsp;turned&nbsp;out&nbsp;to&nbsp;be&nbsp;wrong,&nbsp;so&nbsp;we&nbsp;need&nbsp;to&nbsp;reassembly&nbsp;them</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >7.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Sent&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032,&nbsp;to&nbsp;sequ</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >e</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >nc</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >e</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >8.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Amplified&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361&nbsp;through&nbsp;PCR</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >8.25~8.31</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; color:rgb(40,40,40); font-size:10.5000pt; font-family:'Times New Roman'; " >1.&nbsp;Digested&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032&nbsp;and&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_R0062&nbsp;+&nbsp;BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Designed&nbsp;primer&nbsp;of&nbsp;APO</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Failed&nbsp;to&nbsp;PCR&nbsp;Cre</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ssrA</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Transformed&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >PO,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >5.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;R</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ecultured&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_R0062&nbsp;+&nbsp;BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Single</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >digested&nbsp;BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >o,&nbsp;compared&nbsp;with&nbsp;previous&nbsp;single&nbsp;digested&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;but&nbsp;failed</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >7.&nbsp;Redigested&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;then&nbsp;did&nbsp;ligation&nbsp;and&nbsp;transformation</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >8.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Sen</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >t</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;to&nbsp;sequenc</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >e</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >:&nbsp;&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_R0062&nbsp;+&nbsp;BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >o,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >9.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Recovered&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;and&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040&nbsp;from&nbsp;conservation</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >9.1~9.7</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.&nbsp;succeeded&nbsp;in&nbsp;amplifying&nbsp;Cre</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ssrA&nbsp;by&nbsp;PCR</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Digested&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040,&nbsp;pSB1C3,&nbsp;pSB1K3,&nbsp;pSB1A3,&nbsp;pSB1T3,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_R0062&nbsp;+&nbsp;BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;Cre</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;PCRed&nbsp;BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0015&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >A</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ssembled</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_I718008&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(pSB1C3),</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(pSB1T3),&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >po</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(pSB1T3),</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >RBS</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_I718008&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ssrA</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(pSB1C3)</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0015&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(pSB1C3)</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_I13504&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(pSB1C3),&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_I13504&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(pSB1C3),</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_B0015&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >(pSB1C3),&nbsp;then&nbsp;transformed&nbsp;them</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.&nbsp;Cultured&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_R0062&nbsp;+&nbsp;BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >5.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Di</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >g</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ested&nbsp;pSB1C3,&nbsp;pSB1K3,&nbsp;pSB1A3,&nbsp;pSB1T3,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_R0062&nbsp;+&nbsp;BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_CO261</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;Transformed</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;Cre</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;pSB1A2,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J04650</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >7.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;Transformed</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;pSB1A2,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J04650</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >8.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Sen</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >t</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;to&nbsp;sequenc</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >e</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >:&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_R0062&nbsp;+&nbsp;BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0015&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >B0034</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >9.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Did&nbsp;gel&nbsp;extraction&nbsp;RBS</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Cre</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ssrA</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >9.8~9.14</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Digested&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >pSB1C3</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >o,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >po</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Incubated&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >Assembled</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >po</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I13507,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_J04650,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >po</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Sent&nbsp;to&nbsp;sequenc</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >e</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >:&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >B0034</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;PO&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >5</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >.96-well&nbsp;plates&nbsp;experiment&nbsp;of&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;PO</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >6</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Cultured&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_I13504</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;and&nbsp;observed&nbsp;its&nbsp;fluorescence</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >7</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;Transformed</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_J04650,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >o</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >9.15~9.21</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >1.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;Miniprep:&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I13507,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_J04650,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Po</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I13507,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I13507,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_K145215</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_J04650,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BBA_F2622</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0015&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >o</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_I13521,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J04650&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >2.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;D</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >igestion:</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015,&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J04650,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Po</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >o</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_K145215</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >3.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;Incubate</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >:&nbsp;&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_R0040&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_I718008&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Po</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J04650,&nbsp;BB</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >a</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >_K145215</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J04650,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0034&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Po</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >repared&nbsp;A</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; vertical-align:super; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; vertical-align:super; " >&nbsp;&nbsp;&nbsp;&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >and&nbsp;T</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; vertical-align:super; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; vertical-align:super; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;plate</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >s</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >5.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >Assembled</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215&nbsp;and&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >then&nbsp;did&nbsp;transformation</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >F</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >unction&nbsp;test:&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J04650,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J04650,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J04650</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13521,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507,</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Po</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >7.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >G</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >el&nbsp;extraction&nbsp;of</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;P</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >o</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >8.&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >S</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >ent&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J23032</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >J04650</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215,&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >R0040&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;to&nbsp;sequen</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >ce&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >9.22-9.28</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >1</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Prepare&nbsp;LB&nbsp;incubate&nbsp;and&nbsp;LB&nbsp;plate</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >s</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >2</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Incubated&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0034</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >,&nbsp;and&nbsp;then&nbsp;extracted&nbsp;plasmid&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >4.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Digested&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13521</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >,&nbsp;BBa_I718008,&nbsp;BBa_B0034</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >5.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;G</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >el&nbsp;extraction&nbsp;of&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_I718008,&nbsp;and&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >6.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Function&nbsp;test&nbsp;of</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13521</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >,&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015,&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >K145215</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >,&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I746361</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;+&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >7.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >PCR</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;BBa_</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >I13507</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >8.</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >Assembled&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_I718008</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&nbsp;and&nbsp;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0034</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " >&#8594;</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'&#23435;&#20307;'; " >BBa_B0015</span><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p></o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p><p class=p0  style="margin-bottom:0pt; margin-top:0pt; " ><span style="mso-spacerun:'yes'; font-size:10.5000pt; font-family:'Times New Roman'; " ><o:p>&nbsp;</o:p></span></p></div><!--EndFragment--></body></html>