"
Team:EPF-Lausanne/Notebook/Cloning Strategy
From 2009.igem.org
Contents |
Cloning strategy
21.07.09
One useful website to know the restriction sites of enzymes:
http://www.genscript.com/cgi-bin/products/enzyme.cgi?op=all_ez
The restriction site used were:
EcoRI GAATTC
XbaI TCTAGA
SpeI ACTAGT
PsiI TTATAA
Design the primers for the 2 step-PCR: the first step introduces the LacI promoter and the RBS upstream the LovTAP gene with the Forward primer, whereas the Reverse introduces the Term downstream. In the second step we only introduce the E-X restriction sites upstream and the SP downstream.
