Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/12/15

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Overview

Phusion PCR of HPK:CFP part.

Materials/Methods

Reagent Stock Volume (µL)
GC Buffer5x10
dNTP mix10 mM1
DMSO-1.5
Phusion20 U/µL0.5
Template50 ng0.3
Primer (for)10 µM2
Primer (rev)10 µM2
H2O-32.7
Total50

Template: DBN007
Primers: P147, P151

PCR protocol: basic Phusion, 60°C annealing temp, 90 second extension time, 35 cycles.

Gel Results

My PCR product is on the far right. Lots of non-specific binding and amplifcation. Main bands are at ~1500bp and ~2000bp. Expected product size is 2000bp.


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