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PCR-AFLP - (Mar/02/2005 )

I am planning to do PCR-AFLP. Do I need to precipitate ligated DNA to remove unbound adapters which may serve as primers in the subsequent PCR? thanks for suggestions.

-orhan-

I don't think you have to precipitate the reaction. I have done AFLPs for several years and it always worked without.

-sandrawrobel-

thanks!

-orhan-

also, is there a need for inactivating DNA ligase in water bath after restiriction-ligation step? thanks.

-orhan-

I've never heat-inactivated either the enzymes from the restriction or the ligase. Though I remember trying this when I had problems with the whole AFLP procedure once. It works without, but it wouldn't do harm either, I guess.

-sandrawrobel-

great!

-orhan-