SCU Weekly

From 2013.igem.org

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=='''Weekly Notes'''==
 
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===<center>June</center>===
 
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----
<html>
<html>
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<body>
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6.8~6.15
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<map name="MapMap">
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1. Transformation of BBa_B0015, BBa_CO261 to psb1AK3 is successful
-
  <area shape="rect" coords="1,65,234,92" href="https://2013.igem.org/SCU_Weekly#Week2">
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2. Transformed pSB1T3, pSB1K3, pSB1C3, pSB1A3, BBa_I13504 and BBa_I13504.
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</map>
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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).
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<div style="text-align:center"><img src="https://static.igem.org/mediawiki/2013/0/0b/SCU-Jun.png" width="236" height="159" border="0" align="middle" usemap="#Map"> </div>
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5. Chloramphenicol was insoluble，and it was tested to be useless using DH5α.
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<map name="Map"><area shape="rect" coords="0,137,234,159" href="https://2013.igem.org/SCU_Weekly#Week_5_A"><area shape="rect" coords="0,115,234,137" href="https://2013.igem.org/SCU_Weekly#Week_4"><area shape="rect" coords="1,92,235,114" href="https://2013.igem.org/SCU_Weekly#Week_3"><area shape="rect" coords="6,160,239,187" href="https://2013.igem.org/SCU_Weekly#Week_3">
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6. Prepare A + , K + ,T + ,C +  plates and antibiotic store liquor.
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  <area shape="rect" coords="0,43,234,65" href="https://2013.igem.org/SCU_Weekly#Week_1_and_Week_2">
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7. Digested and assembled BBa_CO261, pSB1C3, then transformed them
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  <area shape="rect" coords="1,65,234,92" href="https://2013.igem.org/SCU_Weekly#Week_1_and_Week_2">
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9. Digestion of BBa_I13504 and BBa_B0015, and results proved that the transformation was successful
-
</map>
+
-
</body>
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-
</html>
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-
===Week 1 and Week 2===
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-
===Week 3===
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-
===Week 4===
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-
===Week 5 <span style="color:transparent"> A</span>===
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-
<div style="text-align:right"> [[#June|<small>'''[June]'''</small>]] [[#July|<small>'''[July]'''</small>]] [[#August|<small>'''[August]'''</small>]] [[#September|<small>'''[September]'''</small>]] [https://2013.igem.org/Team:SCU_China/Notebook <small>'''*To the Protocol Page'''</small>]</div>
+
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
-
===<center>July</center>===
+
6.23~6.29
-
----
+
1. Strain conservation of BBa_F2622 + BBa_CO261, pSB1C3
-
<html>
+
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
-
<body>
+
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
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<div style="text-align:center"> <p><img src="https://static.igem.org/mediawiki/2013/f/f8/SCU-July.png" width="236" height="156" border="0" usemap="#Map1"></div>
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7.14~7.20
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  <map name="Map1"><area shape="rect" coords="7,131,233,152" href="https://2013.igem.org/SCU_Weekly#Week_9_A"><area shape="rect" coords="4,110,230,131" href="https://2013.igem.org/SCU_Weekly#Week_8"><area shape="rect" coords="3,89,229,110" href="https://2013.igem.org/SCU_Weekly#Week_7"><area shape="rect" coords="4,67,230,88" href="https://2013.igem.org/SCU_Weekly#Week_6">
+
1. No much experiment
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    <area shape="rect" coords="5,44,231,65" href="https://2013.igem.org/SCU_Weekly#Week_5_B">
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-
  </map>
+
-
</p>
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-
</body>
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-
</html>
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-
===Week 5 <span style="color:transparent">B</span>===
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7.21~7.27
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===Week 6===
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1. Failed to amplify pSB1K3, pSB1A3, pSB1C3 through PCR
-
===Week 7===
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2. Transformed BBa_I61046 (RBS + Cre), K5-R0040 and they were successful
-
===Week 8===
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3. Extracted plasmid of pSB1T3, BBA_F2622 + BBa_CO261 and only pSB1T3was successful
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===Week 9 <span style="color:transparent">A</span>===
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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:
 +
Taq is suitable for amplifying snippets of 2kb in size, and the reaction is fast
 +
10X taq buffer with (NH4)2SO4 reduces nonspecification
 +
2mM 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.
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<div style="text-align:right"> [[#June|<small>'''[June]'''</small>]] [[#July|<small>'''[July]'''</small>]] [[#August|<small>'''[August]'''</small>]] [[#September|<small>'''[September]'''</small>]] [https://2013.igem.org/Team:SCU_China/Notebook <small>'''*To the Protocol Page'''</small>]</div>
+
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
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===<center>August</center>===
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8.3~8.10
-
----
+
-
<html>
+
 +
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
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<body>
+
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.
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<div style="text-align:center"><p><img src="https://static.igem.org/mediawiki/2013/4/4e/SCU-Aug.png" width="236" height="156" border="0" usemap="#Map2"></div>
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8.18~8.24
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  <map name="Map2"><area shape="rect" coords="6,130,232,151" href="https://2013.igem.org/SCU_Weekly#Week_13"><area shape="rect" coords="4,110,230,131" href="https://2013.igem.org/SCU_Weekly#Week_12"><area shape="rect" coords="3,89,229,110" href="https://2013.igem.org/SCU_Weekly#Week_11"><area shape="rect" coords="4,67,230,88" href="https://2013.igem.org/SCU_Weekly#Week_10">
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1. Assembled BBa_R0040 + BBa_J23032 (Ptet + lock), PO + LOX + BBa_R0040
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    <area shape="rect" coords="5,44,231,65" href="https://2013.igem.org/SCU_Weekly#Week_9_B">
+
2. Transformed BBa_R0062 + BBa_I718008, BBa_F2622 + BBa_CO261, BBa_R0040 + BBa_J23032
-
  </map>
+
3. Incubated BBa_R0040 + BBa_J23032, BBa_R0062 + BBa_I718008
-
</p>
+
4. Extracted plasmid of BBa_J23032, B0034（strong RBS, BBa_R0040 + BBa_J23032, BBa_R0062 + Cre and then did digestion.
-
</body>
+
5. Digested BBa_I718008, BBa_R0062 + BBa_I718008, BBa_B0034.
-
</html>
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6. Sent BBa_R0062 + BBa_I718008to sequence, however all turned out to be wrong, so we need to reassembly them.
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===Week 9 <span style="color:transparent">B</span>===
+
7. Sent BBa_R0040 + BBa_J23032, to sequence
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===Week 10===
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8. Amplified BBa_I746361 through PCR
-
===Week 11===
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-
===Week 12===
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-
===Week 13===
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-
 
+
-
 
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-
 
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<div style="text-align:right"> [[#June|<small>'''[June]'''</small>]] [[#July|<small>'''[July]'''</small>]] [[#August|<small>'''[August]'''</small>]] [[#September|<small>'''[September]'''</small>]] [https://2013.igem.org/Team:SCU_China/Notebook  <small>'''*To the Protocol Page'''</small>]</div>
+
-
 
+
-
===<center>September</center>===
+
-
----
+
-
<html>
+
 +
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
-
<body>
+
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
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<div style="text-align:center"><p><img src="https://static.igem.org/mediawiki/2013/2/21/SCU-Sep1.png" width="236" height="156" border="0" usemap="#Map3"></div>
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9.8~9.14
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  <map name="Map3"><area shape="rect" coords="6,130,232,151" href="https://2013.igem.org/SCU_Weekly#Week_18"><area shape="rect" coords="4,110,230,131" href="https://2013.igem.org/SCU_Weekly#Week_17"><area shape="rect" coords="3,89,229,110" href="https://2013.igem.org/SCU_Weekly#Week_16"><area shape="rect" coords="5,67,231,88" href="https://2013.igem.org/SCU_Weekly#Week_15">
+
1. Digested BBa_I13507, pSB1C3, BBa_R0040 + Po, BBa_R0040 + po + BBa_I13507
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    <area shape="rect" coords="5,44,231,65" href="https://2013.igem.org/SCU_Weekly#Week_14">
+
2. Incubated BBa_R0040→BBa_B0015
-
  </map>
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3. Assembled BBa_R0040 + po + BBa_I13507, BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040 + po + BBa_I13507
-
</p>
+
4. Sent to sequence: BBa_B0034→BBa_B0015, BBa_R0040→BBa_B0015, BBa_R0040 + PO + BBa_I13507
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</body>
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5.96-well plates experiment of BBa_R0040 + PO
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</html>
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6. Cultured BBa_R0040 + BBa_I13504 and observed its fluorescence
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===Week 14===
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7. Transformed BBa_R0040 + BBa_J23032 + BBa_J04650, BBa_R0040 + Po + BBa_I13507
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===Week 15===
+
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===Week 16===
+
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===Week 17===
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===Week 18===
+
 +
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
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<div style="text-align:right"> [[#June|<small>'''[June]'''</small>]] [[#July|<small>'''[July]'''</small>]] [[#August|<small>'''[August]'''</small>]] [[#September|<small>'''[September]'''</small>]] [https://2013.igem.org/Team:SCU_China/Notebook  <small>'''*To the Protocol Page'''</small>]</div>
+
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

Revision as of 03:50, 28 September 2013

Animation


 

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: Taq is suitable for amplifying snippets of 2kb in size, and the reaction is fast 10X taq buffer with (NH4)2SO4 reduces nonspecification 2mM Mg2 + is suitable for taq Avoid DNA pollution during template preparation No pollution of raw material, it would be better to operate on the laminar Negative control is necessary. 7.28~8.2 1. Recovered PCR product of 27th 2. 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 |} __NOEDITSECTION__ __NOTOC__  
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