Team:UNITN-Trento/Notebook/Labposts/08/06

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(Created page with "{ "date" : "2013-08-05", "author" : "thomas", "title" : "EFE PCR", "content" : "<html>Today I performed a PCR on EFE in pUC57 vector in order to linearize it and eliminate th...")

Revision as of 07:29, 9 September 2013

{ "date" : "2013-08-05", "author" : "thomas", "title" : "EFE PCR", "content" : "Today I performed a PCR on EFE in pUC57 vector in order to linearize it and eliminate the vector. To do this I used primers BBa_Fwd and BBa_Rev. The reaction was assembled as follow exploting both OneTaq and Phusion polymerase.

5x One Taq Buffer10µl
Fwd Primer1µl
Rev Primer1µl
10 mM dNTP's1µl
One Taq0.25µl
Phusion0.3µl
Template DNA0.7µl (about 500 ng)
H2O35.75µl
When the reaction finished, I performed an electrophoresis analysis in order to confirm the product. Kapa universal ladder was adopted.
Gel image
As you can see, the Gel image is quite confusing. First of all, the marker should be diffused in the sample lane (lane 1) as we can see the same band of the ladder but with a more tenuous coloration. Secondly, the thickest band was at about 1200 bp height even if EFE is 1078 bp long. I think the product has to be confirmed again...!", "tags" : "EFE" }

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