Plasmids 101: Methylation and Restriction Enzymes
Type
Blog Post
Published
June 30, 2016, 2:30 p.m.
...desired mutation into your plasmid of interest by PCR generates mutated plasmids with no methylation (there...Mutagenesis to Mutate Your Plasmids
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Boost Your Molecular... for purposes other than cloning or confirming plasmids. It turns out that restriction enzymes are one...difficult to interpret or lead you to conclude your plasmid is not correct!
Conversely, enzymes such as DpnI...methyltransferases in the PCR reaction). The template plasmid, on the other hand, should be derived from a dam...methylated adenines in any GATC sequences found in the plasmid. When the PCR products are digested with DpnI, ... is destroyed leaving behind a pool of mutated plasmids which can later be verified by Sanger sequencing...