Plasmids undergoing NHEJ?
I am trying to screen for gRNA's targeting a plasmid that is being co-transfected into HEK 293T cells. However, when I sequence around the cut site for TIDE, I am not seeing any detectable indel formations.
Has anyone done this before? Can plasmids undergo NHEJ, or is it that the plasmids undergo degradation after cutting?
Kevin
I've done this before. We noticed that you can do a T7E assay, but the yield was highly variable (and even from the same sample it was PCR dependent) and it seemed like a large portion of the plasmid was being degraded instead of repaired.
Michael