Hi guys,

I tried a CRIRPS mediated knockout on HEK-cells and unfortunately it wasn't successful.

Any ideas what could be the reason?

There are several points which would come to my mind, like CRISPR efficiency on a tetraploid organism?

I only did one round of transfection (and did not check transfection-efficiency with GFP)

I know for sure that the plasmid had the correct gRNA build in (sent it away for sequencing)

Another point could maybe be the selection with puromycin?

Anybody got any suggestions?

(I am a newby in serios lab-stuff)

thanks and best regards!