SCU Weekly

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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 (red) - 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 assembled BBa_CO261, pSB1C3, then transformed them

9. Digestion of BBa_I13504 and BBa_B0015, and results proved that the transformation was successful

6.16~6.22

1. PCR to amplify 8B but failed.

2. Chloramphenicol was tested to be effective

3. Transformed pSB1T3, BBa_CO261, BBa_B0015, BBa_I13504, pSB1K3 (pSB1K3) and pSB1A3(pSB1A3)

4. Plasmid extraction of BBa_F2622 + BBa_CO261

5. Digested BBa_F2622 + BBa_CO261 and pSB1T3 ( EcoRⅠand PstⅠ)

6. Assembled BBa_F2622 + BBa_CO261, BBa_B0015 and pSB1T3

6.23~6.29

1. Strain conservation of BBa_F2622 + BBa_CO261, pSB1C3

2. Prepared ddH2O

3. Added 1µl Dpn Ӏ into pSB1T3 (pSB1T3) reaction system when digesting

6.30~7.6

1. Digested BBa_F2622 + BBa_CO261 and pSB1T3 ( EcoRⅠand PstⅠ)

2. Assembled BBa_F2622 + BBa_CO261, BBa_I13504, and pSB1T3, but failed to transform them

7.7~7.13

1. Plated BBA_F2622 + BBa_CO261 + BBa_I13504 (1.2.3) and BBA_F2622 + BBa_CO261 + BBa_B0015 (1.2.3) which were assembled 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 BBa_I61046 (RBS + Cre), K5-R0040 and they were successful

3. Extracted plasmid of pSB1T3, BBA_F2622 + BBa_CO261 and only pSB1T3was successful

4. Incubated BBa_R0040, BBa_F2622 + BBa_CO261 + BBa_I13504, BBA_F2622 + BBa_CO261 + BBa_B0015

5. Successfully assembled BBA_F2622 + BBa_CO261 + BBa_I13504, BBA_F2622 + BBa_CO261 + BBa_B0015

RePCR by taq enzyme, we got experiences:

1. Taq is suitable for amplifying snippets of 2kb in size, and the reaction is fast

2. 10X taq buffer with (NH4)2SO4 reduces nonspecification

3. 2mM Mg2 + is suitable for taq

4. Avoid DNA pollution during template preparation

5. No pollution of raw material, it would be better to operate on the laminar

6. Negative control is necessary.

7.28~8.2

1. Recovered PCR product of 27th

2. Assembled BBa_F2622 + BBa_CO261 + BBa_I13504, BBa_F2622 + BBa_CO261 + BBa_B0015 with pSB1C3 carrier, then did transformation and incubate

3. Incubated R0040, pSB1T3, BBa_R0062, BBa_F2622 + BBa_CO261, BBa_B0015

4. Replated BBa_F2622 + BBa_CO261 + BBa_I13504, BBa_F2622 + BBa_CO261 + BBa_B0015

5. Digested BBa_R0040, BBa_R0062, BBa_J61064

6. Assembled BBa_R0062with BBa_J61064, then did transformation

8.3~8.10

1. Incubated BBa_R0062+BBa_J61064, and gel analysis of plasmid extraction and digestion products proved that the ligation was successful

2. Transformed BBa_I746361, BBa_K145215, BBa_K145215.

3. Digested BBa_R0062, BBa_J61064, pSB1C3, and assembled them overnight

8.11~8.17

1. Transformed, BBa_I746361, BBa_K145215，BBa_J23032，BBa_F2622 + BBa_CO261

2. Extracted plasmid of BBa_K145215, BBa_F2622 + BBa_CO261

3. Digested BBa_K145215, BBa_CO261, BBa_F2622 + BBa_CO261，

4. Incubated BBa_F2622，BBa_J23032

5. Sent BBa_K145215, BBA_F2622 to biotech company to sequence them.

8.18~8.24

1. Assembled BBa_R0040 + BBa_J23032 (Ptet + lock), PO + LOX + BBa_R0040

2. Transformed BBa_R0062 + BBa_I718008, BBa_F2622 + BBa_CO261, BBa_R0040 + BBa_J23032

3. Incubated BBa_R0040 + BBa_J23032, BBa_R0062 + BBa_I718008

4. Extracted plasmid of BBa_J23032, B0034（strong RBS, BBa_R0040 + BBa_J23032, BBa_R0062 + Cre and then did digestion.

5. Digested BBa_I718008, BBa_R0062 + BBa_I718008, BBa_B0034.

6. Sent BBa_R0062 + BBa_I718008to sequence, however all turned out to be wrong, so we need to reassembly them.

7. Sent BBa_R0040 + BBa_J23032, to sequence

8. Amplified BBa_I746361 through PCR

8.25~8.31

1. Digested BBa_R0040 + BBa_J23032 and BBa_F2622 + BBa_B0015, BBa_R0062 + BBa_I718008

2. Designed primer of APO

3. Failed to PCR Cre + ssrA

4. Transformed BBa_R0040 + PO,

5. Recultured BBa_R0062 + BBa_I718008, BBA_F2622 + BBa_CO261

6. Single digested BBA_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: BBa_R0040 + BBa_J23032, BBa_R0062 + BBa_I718008, R0040 + Po,

9. Recovered BBa_B0015 and BBa_R0040 from conservation

9.1~9.7

1. succeeded in amplifying Cre + ssrA by PCR

2. Digested BBa_R0040, pSB1C3, pSB1K3, pSB1A3, pSB1T3, BBa_R0062 + BBa_I718008, BBA_F2622 + BBa_CO261, Cre + BBa_B0015, PCRed BBA_F2622 + BBa_B0015, BBA_F2622 + BBa_B0015 + R0040 + J23032

3.Assembled BBa_I718008 + BBa_B0015, BBa_F2622 + BBa_CO261(pSB1C3), R0040 + J23032 + BBA_F2622 + BBa_B0015(pSB1T3), BBa_R0040 + po + BBa_I13507 (pSB1T3), RBS + BBa_I718008 + ssrA (pSB1C3) ,BBa_F2622 + BBa_B0015 (pSB1C3) BBa_R0040 + BBa_I13504 (pSB1C3), BBa_I746361 + BBa_I13504 (pSB1C3), BBa_R0040 → BBa_B0015 (pSB1C3), then transformed them.

4. Cultured BBa_R0062 + BBa_I718008

5. Digested pSB1C3, pSB1K3, pSB1A3, pSB1T3, BBa_R0062 + BBa_I718008, BBa_F2622 + BBa_CO261

6. Transformed Cre + BBa_B0015, pSB1A2, BBa_J04650

7. Transformed pSB1A2, BBa_J04650

8. Sent to sequence: BBa_R0062 + BBa_I718008, BBa_F2622 + BBa_B0015 → 1B0034

9. Did gel extraction RBS + Cre + ssrA

9.8~9.14

1. Digested BBa_I13507, pSB1C3, BBa_R0040 + Po, BBa_R0040 + po + BBa_I13507

2. Incubated BBa_R0040→BBa_B0015

3. Assembled BBa_R0040 + po + BBa_I13507, BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040 + po + BBa_I13507

4. Sent to sequence: BBa_B0034→BBa_B0015, BBa_R0040→BBa_B0015, BBa_R0040 + PO + BBa_I13507

5.96-well plates experiment of BBa_R0040 + PO

6. Cultured BBa_R0040 + BBa_I13504 and observed its fluorescence

7. Transformed BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040 + Po + BBa_I13507

9.15~9.21

1. Miniprep: BBa_I746361 + BBa_I13507, BBa_R0040 + BBa_ J23032 + BBa_J04650, BBa_R0040 + Po + BBa_I13507, BBa_I746361 + BBa_I13507, BBa_I746361 + BBa_I13507 + BBa_R0040→BBa_B0015, BBa_K145215 + BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_I746361 + BBa_I13507 + BBa_R0040 + BBa_J23032 + BBA_F2622 + BBa_B0015 + BBa_K145215, BBa_R0040 + Po + BBa_I13507, BBa_I746361→BBa_B0015 + BBa_K145215, BBa_I13521, BBa_K145215 + BBa_R0040 + BBa_J23032 + BBa_J04650

2. Digestion: BBa_I746361 + BBa_I13507, BBa_I746361 + BBa_I13507 + BBa_R0040→BBa_B0015, BBa_K145215 + BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040 + Po + BBa_I13507, BBa_R0040 + Po + BBa_I13507, BBa_I746361→BBa_B0015 + BBa_K145215

3. Incubate: BBa_I746361 + BBa_I13507, BBa_I746361 + BBa_I13507 + BBa_R0040→BBa_B0015, BBa_I718008 + Po + BBa_I13507, BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_K145215 + BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040, BBa_K145215 + BBa_B0034 + BBa_J23032 + BBa_B0015, R0040 + Po + BBa_I13507

4. Prepared A + and T + plates

5. Assembled BBa_I746361→BBa_B0015 + BBa_K145215 and BBa_R0040 + BBa_I13507, then did transformation

6. Function test: BBa_K145215 + BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040 + BBa_J23032 + BBa_J04650 + BBa_K145215, BBa_I13521, BBa_I746361 + BBa_I13507, BBa_I746361 + BBa_I13507 + BBa_R0040→BBa_B0015, BBa_R0040 + Po + BBa_I13507

7. Gel extraction of BBa_R0040 + Po + BBa_I13507

8. Sent BBa_R0040 + BBa_J23032 + BBa_J04650 + BBa_K145215, BBa_I746361 + BBa_I13507 + BBa_R0040→BBa_B0015 to sequence

9.22-9.28

1. Prepare LB incubate and LB plates

2. Incubated BBa_B0034→BBa_B0015, and then extracted plasmid

4. Digested BBa_I13521, BBa_I718008, BBa_B0034→BBa_B00155. Gel extraction of BBa_I718008, and BBa_I13507

6. Function test of BBa_I13521, BBa_I746361→BBa_B0015, BBa_I746361→ BBa_K145215, BBa_I746361 + BBa_I13507

7. PCR BBa_I13507

8. Assembled BBa_I718008 and BBa_B0034→BBa_B0015

@@ Line 27: / Line 27: @@
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-.8~6.15
+@font-face{
-. Transformation of BBa_B0015, BBa_CO261 to psb1AK3 is successful
+font-family:"&#24494;&#36719;&#38597;&#40657;";
-. 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 (red) - 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 assembled BBa_CO261, pSB1C3, then transformed them
-. Digestion of BBa_I13504 and BBa_B0015, and results proved that the transformation was successful
-.16~6.22
+@font-face{
-. PCR to amplify 8B but failed.
+font-family:"Tahoma";
-. Chloramphenicol was tested to be effective
+}
-. Transformed pSB1T3, BBa_CO261, BBa_B0015, BBa_I13504, pSB1K3 (pSB1K3) and pSB1A3(pSB1A3)
-. Plasmid extraction of BBa_F2622 + BBa_CO261
-. Digested BBa_F2622 + BBa_CO261 and pSB1T3 ( EcoRⅠand  PstⅠ)
-. Assembled BBa_F2622 + BBa_CO261, BBa_B0015 and pSB1T3
-.23~6.29
+@font-face{
-. Strain conservation of BBa_F2622 + BBa_CO261, pSB1C3
+font-family:"Symbol";
-. Prepared ddH2O
+}
-. Added 1µl Dpn Ӏ into pSB1T3 (pSB1T3) reaction system when digesting
-.30~7.6
+@font-face{
-. Digested BBa_F2622 + BBa_CO261 and pSB1T3 ( EcoRⅠand PstⅠ)
+font-family:"Arial";
-. Assembled BBa_F2622 + BBa_CO261, BBa_I13504, and pSB1T3, but failed to transform them
+}
-.7~7.13
+@font-face{
-. Plated BBA_F2622 + BBa_CO261 + BBa_I13504 (1.2.3) and BBA_F2622 + BBa_CO261 + BBa_B0015 (1.2.3) which were assembled last week but failed.
+font-family:"&#40657;&#20307;";
-. Cultured BBa_B0015 1 and 2
+}
-.14~7.20
+@font-face{
-. No much experiment
+font-family:"Courier New";
+}
-.21~7.27
+@font-face{
-. Failed to amplify pSB1K3, pSB1A3, pSB1C3 through PCR
+font-family:"Wingdings";
-. Transformed BBa_I61046 (RBS + Cre), K5-R0040 and they were successful
+}
-. Extracted plasmid of pSB1T3, BBA_F2622 + BBa_CO261 and only pSB1T3was successful
-. Incubated BBa_R0040, BBa_F2622 + BBa_CO261 + BBa_I13504, BBA_F2622 + BBa_CO261 + BBa_B0015
-. Successfully assembled BBA_F2622 + BBa_CO261 + BBa_I13504, BBA_F2622 + BBa_CO261 + BBa_B0015
-RePCR by taq enzyme, we got experiences:
+@font-face{
-Taq is suitable for amplifying snippets of 2kb in size, and the reaction is fast
+font-family:"Calibri";
-X taq buffer with (NH4)2SO4 reduces nonspecification
+}
-mM 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.
-.28~8.2
+@font-face{
-. Recovered  PCR product of 27th
+font-family:"Cambria Math";
-. Assembled BBa_F2622 + BBa_CO261 + BBa_I13504, BBa_F2622 + BBa_CO261 + BBa_B0015 with pSB1C3 carrier, then did transformation and incubate
+}
-. Incubated R0040, pSB1T3, BBa_R0062, BBa_F2622 + BBa_CO261, BBa_B0015
-. Replated BBa_F2622 + BBa_CO261 + BBa_I13504, BBa_F2622 + BBa_CO261 + BBa_B0015
-. Digested BBa_R0040, BBa_R0062, BBa_J61064
-. Assembled BBa_R0062with BBa_J61064, then did transformation
-.3~8.10
+p.p0{
+margin:0pt;
+margin-bottom:0.0001pt;
+margin-bottom:0pt;
+margin-top:0pt;
+text-align:justify;
+font-size:10.5000pt; font-family:'Calibri'; }
-. Incubated BBa_R0062+BBa_J61064, and gel analysis of plasmid extraction and digestion products proved that the ligation was successful
+span.10{
-. Transformed BBa_I746361, BBa_K145215, BBa_K145215.
+font-size:10.0000pt; font-family:'Times New Roman'; }
-. Digested BBa_R0062, BBa_J61064, pSB1C3, and assembled them overnight
-.11~8.17
+span.15{
-. Transformed, BBa_I746361, BBa_K145215，BBa_J23032，BBa_F2622 + BBa_CO261
+font-size:10.0000pt; font-family:'Times New Roman'; }
-. Extracted plasmid of BBa_K145215, BBa_F2622 + BBa_CO261
-. Digested BBa_K145215, BBa_CO261, BBa_F2622 + BBa_CO261，
-. Incubated BBa_F2622，BBa_J23032
-. Sent BBa_K145215, BBA_F2622 to biotech company to sequence them.
-.18~8.24
+span.16{
-. Assembled BBa_R0040 + BBa_J23032 (Ptet + lock), PO + LOX + BBa_R0040
+font-size:10.0000pt; font-family:'Times New Roman'; }
-. Transformed BBa_R0062 + BBa_I718008, BBa_F2622 + BBa_CO261, BBa_R0040 + BBa_J23032
-. Incubated BBa_R0040 + BBa_J23032, BBa_R0062 + BBa_I718008
-. Extracted plasmid of BBa_J23032, B0034（strong RBS, BBa_R0040 + BBa_J23032, BBa_R0062 + Cre and then did digestion.
-. Digested BBa_I718008, BBa_R0062 + BBa_I718008, BBa_B0034.
-. Sent BBa_R0062 + BBa_I718008to sequence, however all turned out to be wrong, so we need to reassembly them.
-. Sent BBa_R0040 + BBa_J23032, to sequence
-. Amplified BBa_I746361 through PCR
-.25~8.31
+span.17{
-. Digested BBa_R0040 + BBa_J23032 and BBa_F2622 + BBa_B0015, BBa_R0062 + BBa_I718008
+color:rgb(53,161,212); text-decoration:underline ;font-size:10.0000pt; font-family:'Times New Roman'; }
-. Designed primer of APO
-. Failed to PCR Cre + ssrA
-. Transformed BBa_R0040 + PO,
-. Recultured BBa_R0062 + BBa_I718008, BBA_F2622 + BBa_CO261
-. Single digested BBA_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:  BBa_R0040 + BBa_J23032, BBa_R0062 + BBa_I718008, R0040 + Po,
-. Recovered BBa_B0015 and BBa_R0040 from conservation
-.1~9.7
+span.18{
-. succeeded in amplifying Cre + ssrA by PCR
+font-size:9.0000pt; font-family:'&#23435;&#20307;'; }
-. Digested BBa_R0040, pSB1C3, pSB1K3, pSB1A3, pSB1T3, BBa_R0062 + BBa_I718008, BBA_F2622 + BBa_CO261, Cre + BBa_B0015, PCRed BBA_F2622 + BBa_B0015, BBA_F2622 + BBa_B0015 + R0040 + J23032
-.Assembled BBa_I718008 + BBa_B0015, BBa_F2622 + BBa_CO261(pSB1C3), R0040 + J23032 + BBA_F2622 + BBa_B0015(pSB1T3), BBa_R0040 + po + BBa_I13507 (pSB1T3), RBS + BBa_I718008 + ssrA (pSB1C3) ,BBa_F2622 + BBa_B0015 (pSB1C3) BBa_R0040 + BBa_I13504 (pSB1C3), BBa_I746361 + BBa_I13504 (pSB1C3),  BBa_R0040 → BBa_B0015 (pSB1C3), then transformed them.
-. Cultured BBa_R0062 + BBa_I718008
-. Digested pSB1C3, pSB1K3, pSB1A3, pSB1T3, BBa_R0062 + BBa_I718008, BBa_F2622 + BBa_CO261
-. Transformed Cre + BBa_B0015, pSB1A2, BBa_J04650
-. Transformed pSB1A2, BBa_J04650
-. Sent to sequence: BBa_R0062 + BBa_I718008, BBa_F2622 + BBa_B0015 → 1B0034
-. Did gel extraction RBS + Cre + ssrA
-.8~9.14
+span.19{
-. Digested BBa_I13507, pSB1C3, BBa_R0040 + Po, BBa_R0040 + po + BBa_I13507
+font-size:10.0000pt; font-family:'Times New Roman'; }
-. Incubated BBa_R0040→BBa_B0015
-. Assembled BBa_R0040 + po + BBa_I13507, BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040 + po + BBa_I13507
-. Sent to sequence: BBa_B0034→BBa_B0015, BBa_R0040→BBa_B0015, BBa_R0040 + PO + BBa_I13507
-.96-well plates experiment of BBa_R0040 + PO
-. Cultured BBa_R0040 + BBa_I13504 and observed its fluorescence
-. Transformed BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040 + Po + BBa_I13507
-.15~9.21
+p.p20{
-. Miniprep: BBa_I746361 + BBa_I13507, BBa_R0040 + BBa_ J23032 + BBa_J04650, BBa_R0040 + Po + BBa_I13507, BBa_I746361 + BBa_I13507, BBa_I746361 + BBa_I13507 + BBa_R0040→BBa_B0015, BBa_K145215 + BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_I746361 + BBa_I13507 + BBa_R0040 + BBa_J23032 + BBA_F2622 + BBa_B0015 + BBa_K145215, BBa_R0040 + Po + BBa_I13507, BBa_I746361→BBa_B0015 + BBa_K145215, BBa_I13521, BBa_K145215 + BBa_R0040 + BBa_J23032 + BBa_J04650
+margin-bottom:0pt;
-. Digestion: BBa_I746361 + BBa_I13507, BBa_I746361 + BBa_I13507 + BBa_R0040→BBa_B0015, BBa_K145215 + BBa_R0040 + BBa_J23032 + BBa_J04650,  BBa_R0040 + Po + BBa_I13507, BBa_R0040 + Po + BBa_I13507, BBa_I746361→BBa_B0015 + BBa_K145215
+margin-top:0pt;
-. Incubate:  BBa_I746361 + BBa_I13507, BBa_I746361 + BBa_I13507 + BBa_R0040→BBa_B0015, BBa_I718008 + Po + BBa_I13507, BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_K145215 + BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040, BBa_K145215 + BBa_B0034 + BBa_J23032 + BBa_B0015, R0040 + Po + BBa_I13507
+text-align:justify;
-. Prepared A +     and T +   plates
+font-size:9.0000pt; font-family:'&#23435;&#20307;'; }
-. Assembled BBa_I746361→BBa_B0015 + BBa_K145215 and BBa_R0040 + BBa_I13507, then did transformation
-. Function test: BBa_K145215 + BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040 + BBa_J23032 + BBa_J04650 + BBa_K145215, BBa_I13521, BBa_I746361 + BBa_I13507, BBa_I746361 + BBa_I13507 + BBa_R0040→BBa_B0015, BBa_R0040 + Po + BBa_I13507
-. Gel extraction of BBa_R0040 + Po + BBa_I13507
-. Sent BBa_R0040 + BBa_J23032 + BBa_J04650 + BBa_K145215, BBa_I746361 + BBa_I13507 + BBa_R0040→BBa_B0015 to sequence
-.22-9.28
+p.p21{
-. Prepare LB incubate and LB plates
+margin-bottom:0pt;
-. Incubated BBa_B0034→BBa_B0015, and then extracted plasmid
+margin-top:0pt;
-. Digested BBa_I13521, BBa_I718008, BBa_B0034→BBa_B00155. Gel extraction of BBa_I718008, and BBa_I13507
+text-align:justify;
-. Function test of BBa_I13521, BBa_I746361→BBa_B0015, BBa_I746361→ BBa_K145215, BBa_I746361 + BBa_I13507
+font-size:10.5000pt; font-family:'Calibri'; }
-. PCR BBa_I13507
-. Assembled BBa_I718008 and BBa_B0034→BBa_B0015
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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>