| | Re: Need a cloning genius
No, the overhangs need to be of a particular sequence.
To put into context, I have used Cas9 nickase activity to cleave the genome where I want to create a knock in. The nature of my Cas9 digest has created two 5' overhangs of 55bp. I need to clone 55bp overhangs complementary to the genomic overhang to aid ligation-gated recombination by non-homologous end joining. The sequence cannot be altered as it is within the open reading frame of the gene, and I need it to be expressed.