Stratagene QuikChange mutagenesis?

Troels Wind wind at biobase.dk
Mon Feb 10 05:17:14 EST 1997


Greg (jquinn at nntp.best.com) wrote:
: A question for those who might use this brand of mutagenesis kit:
: I am currently having some problems with a high background using the Stratagene
: QuikChange kit. Does anyone have any experience in reducing this?
: Thanks for any help

This may be stated in the protocol, but anyway...

You can try the following:

	Increase the amount of DpnI in your digestion and/or perform it
	for longer time. This should reduce background.

	Perform several reactions with varying amounts of parental plasmid.
	I do 2, 5, 10 and 25 ng and pick colonies from the reaction with
	the lowest amount. (The lower the amount of parental plasmid, the
	better the chance that that the DpnI digest is complete.)

Good luck!

Troels Wind



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