IGEM:British Columbia/2009/Notebook/Biosensor Sensitivity/2009/08/18
From OpenWetWare
iGEM Project name 1 | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
Plan
Step 1: Anneal lox(R) oligosperformed according to standard IDT oligo kit instructions. Step 2: Addition of LVA tag to Cre through PCRExpected length: 1.2 kb
Forward (FW): cre-lva fw Reverse (RE): cre-LVA re
Gel electrophoresis of PCRed Cre-LVACorrect bands were observed, suggesting that the constructs were successful. Purification of PCR amplified productusing Invitrogen ChargeSwitch kit according to manufacturer's instructions
3. Restriction Digestusing standard UBC iGEM digestion protocol
Cre-LVA and Lox(R)
Incubated reaction mixture for 1hour, 37 degrees Celcius 4. ligation
A. For Lox
B. For Cre-LVA
Transformationperformed using standard UBC iGEM protocol using LB plate with appropriate antibiotics |