Team:UNICAMP-Brazil/Notebooks/September 22

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ColiGuard

Electroelution verification

The samples from yesterday's electroelution were applied to a 0,8% agarose gel and submitted to 80 V during 30 minutes. A band of approximately 22 kb was visible, indicating that the F plasmid was succesfully purified!
Now that the F plasmid was obtained, the next step is its recircularization.

Gabriel

finO and finP - Still Trying to Confirm our Biobricks

We inoculated those colonies that actually resulted in a expected amplification size fragment after the miniprep procedure (recent assays), separately into liquid LB-AMP medium, in order to repeat the miniprep, aiming in reducing the amount of genomic DNA extracted together.

Marcelo

CeaB and CeB: Previous treatment, ligation and transformation

In order to avoid recircularization of BBa_B0015 plasmid, we tried it with alkaline phosphatase (Protocol 9) . After, we started the ligation reaction of this unphosphorylated plasmid with CeaB and CeiB respectively. Considering that this dephosphorylation uses a buffer containing salt and that this salt could be interfering with the transformations, we performed a dyalisis to get the samples out of salts. At last, we transformated competent E. coli (according to the Protocol 3) and plated the respective plates. We expect great results for tomorrow!!!

Luige and Ane

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 ====Electroelution verification====
-*The samples from yesterday's electroelution were applied to a 0,8% agarose gel and submitted to 80 V during 30 minutes. A band of approximately 22 kb was formed, indicating that the F plasmid was succesfully purified!
+*<p style=”text-align:justify;”>The samples from yesterday's electroelution were applied to a 0,8% agarose gel and submitted to 80 V during 30 minutes. A band of approximately 22 kb was visible, indicating that the F plasmid was succesfully purified!</p>
-*Now that the F plasmid was obtained, the next step is its recircularization.
+*<p style=”text-align:justify;”>Now that the F plasmid was obtained, the next step is its recircularization.</p>
 ''Gabriel''
@@ Line 17: / Line 15: @@
 ====finO and finP - Still Trying to Confirm our Biobricks====
-* We innoculated those colonies that actually resulted in a expected amplification size fragment after the miniprep procedure (recent assays), separately into liquid LB-AMP media, in order to repeat the miniprep, aiming in reducing the amount of genomic DNA extracted together.
+*<p style=”text-align:justify;”>We inoculated those colonies that actually resulted in a expected amplification size fragment after the miniprep procedure (recent assays), separately into liquid LB-AMP medium, in order to repeat the miniprep, aiming in reducing the amount of genomic DNA extracted together.</p>
 ''Marcelo''
+====CeaB and CeB: Previous treatment, ligation and transformation====
+*<p style=”text-align:justify;”>In order to avoid recircularization of BBa_B0015 plasmid, we tried it with alkaline phosphatase ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/SAP_Dephosphorylation Protocol 9]) . After, we started the ligation reaction of this unphosphorylated plasmid  with CeaB and CeiB respectively. Considering that this dephosphorylation uses a buffer containing salt and that this salt could be interfering with the transformations, we performed a dyalisis to get the samples out of salts. At last, we transformated competent ''E. coli'' (according to the [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation Protocol 3]) and plated the respective plates.  We expect great results for tomorrow!!!</p>
+''Luige and Ane''
 {{:Team:UNICAMP-Brazil/inc_rodape}}