Restriction digest as a template - (Jun/26/2006 )
Hi, simple question. If I want to use my vector linearised as a template for a subsequent PCR, what should I do? Can I do restriction digestion and then use some ul of the product as a template? Must I purify it before? How? Will any inhibitor be present if I don't do any purification step?
I've tried to find this information unsuccessfully. It's a silly question, I know.
I would clean it. Although you will use a tiny amount for PCR, I would try to get rid of all the salt you have in your digestion
Dilute it 100x, add 1 ul to your PCR reaction and go. No need to linearize or otherwise treat it. Make sure you don't add too much to the PCR reaction.