Restriction digestion of PCR fragments
B VISSER * x2818 Plantkunde *
pbv at rs.uovs.ac.za
Fri Aug 18 08:59:15 EST 1995
Hi
I am in the process of digesting amplified cDNA fragments with Not1 of which
a restriction site was incoporated into the primers. I am not able to fully
digest the fragments, even though sufficient nucleotides are present on the
5' side of the restriction site. Could Ethidium bromide play a role in the
inhibition of the digestion of the DNA? I usually run the fragments on a
prep gel and cut the desired fragments from the gel with phenol/chloroform
which is followed by a ethanol precipitation. Could this be the reason?
Botma Visser
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