How to amplify the insert from a plasmid? - (Aug/03/2005 )
If i am having gene of interest present on Plasmid. And i want to do PCR of that particular gene. How shud i proceed for PCR??? I mean do we hav to get Plasmid linearized prior to PCR or it can work with normal cycles of PCR having profile of Denaturation,annealing and extension.
You don't need to do anything special. Just dilute the plasmid to the appropriate concentration (~ 1 ng total ought to be fine). Standard PCR methods work just fine.
i usually get the same procedure as a standard fragment amplification, without any preprocedure. Except i dilute the plasmid at 10ng/µl and pick 1µl for PCR.