Hi, anyone have suggestions for crispr cleavage detection using thermo genomic cleavage detection kit?
I am trying to make nfkb1 knock out in RAW 264.7 cell line and I have using thermo Cas9 protein system.
After getting intact singe band with transfected cell lysate, i did re-annealing in beaer containin g water at 70 degree and slowing allowing to come 25 degree. I also tried same in water bath.
But in all cases i diidnt got two bands.
even with control template and primers, provided in kit , i am not getting two band.
Can anyone tell me some other protocol for re-annealing step?
Anyone suucceful with thermo GCD kit?