I am looking to perform a double digestion with PacI and NheI. NEB tells me that NheI is affected by some combinations of methylation. My plan is to PCR amplify an ORF from cDNA with restriction site compatible ends and clone it into a plasmid grown up in DH5a cells. How do I know if methylation will affect the NheI digestion on the plasmid?
Edited by BSM, 21 August 2011 - 11:20 AM.