As some of you might be aware I am new to cloning and Crispr/Cas9 gene editing. As a result I've also not carried out many DNA related experiments.
I have recently attempted to make 4 different knock out cell lines using pLentiCrispV2 plasmids. The guides I designed worked for 2 of these cell lines and these were easy to validate as I had good antibodies.
I have terrible antibodies for the other 2 cell lines and have had confusing results with these so I cant tell whether there is a knock out there at all.
I wanted some advice on how to verify these cell lines for a knock out. If I extract DNA from WT and these polyclonal 'KO' cell lines, then PCR amplify a region, can I hand this in for sequencing directly. Considering this is a polyclonal cell line, I'm guessing the results won't be very clean?
Please advise how I can verify these cell lines and the simplest protocol to follow?
Any help would be greatly appreciated.