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?

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