Team:UNICAMP-Brazil/Notebooks/September 4

(Difference between revisions)

Revision as of 01:51, 18 October 2009

Today we prepared electrocompetent E. coli to use in our transformations according to Protocol 4.

Today we started the construction of Cre-Recombinase without ATG, in order to made a new biobrick involving this construction. We ressuspended the biobrick of Cre-Recombinase: BBa_J61047 and transformed in E. coli DH10B eletrocompetent cells. We then plated on LB Amp plate, according to information from registry page.

Víctor

Today we performed the PCR amplification of the PY promoter from the F plasmid previously extracted from the conjugative strain (July 16). We did two different reactions, using two types of forward primers (Ppy-F-1 and Ppy-F-2) for the same reverse primer (Ppy-R). The product amplified with the Ppy-F-1/Ppy-R primers has 133 bp and the one amplified with the Ppy-F-2/Ppy-R has 75 bp. We called these products PY1 and PY2 respectively. The sizes of the products were confirmed by an agarose gel:

Fabiana and Leonardo

@@ Line 18: / Line 18: @@
 ''Víctor''
-==== PY Promoter ====
+==== PY Promoter - PCR amplification ====
 *<p style=”text-align:justify;”>Today we performed the PCR amplification of the PY promoter from the F plasmid previously extracted from the conjugative strain (July 16). We did two different reactions, using two types of forward primers (Ppy-F-1 and Ppy-F-2) for the same reverse primer (Ppy-R). The product amplified with the Ppy-F-1/Ppy-R primers has 133 bp and the one amplified with the Ppy-F-2/Ppy-R has 75 bp. We called these products PY1 and PY2 respectively. The sizes of the products were confirmed by an agarose gel: