Why are PCR products non-methylated ?

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Charvi Nangia

True..but the DNA methyl transferases would methylate them soon...isn't it? How long do they stay unmethylated?

Suzana Radojkovic

Did you also add methyl donor ?

Gertrudis Rojas

In vitro synthesized DNA strands (that have never been inside a cell) remain non-methylated. Template strands coming from E.coli remain methylated.These are the bases for DpnI discrimination.

I understood. Thank you Jan Lennart Von Hacht and Gertrudis Rojas.

@Suzana.. I do not add any methyl donor.

Osama Ghawil

why you studding this ?

Amruta Dhawale

So is it necessary to do DpnI digestion for every PCR products which we want to transform or use in ligation experiments ?