Team:UNICAMP-Brazil/Notebooks/September 22

(Difference between revisions)

Revision as of 02:59, 22 October 2009

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

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

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. In view of this dephosphorylation uses buffer salt that could be interfering in the next transformation, we performed a dyalisis so that get samples free 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 & Ane

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 ====Electroelution verification====
-*<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 formed, indicating that the F plasmid was succesfully purified!</p>
+*<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>
 *<p style=”text-align:justify;”>Now that the F plasmid was obtained, the next step is its recircularization.</p>