发信人: netbuggie (中原一点虫), 信区: Biology
标 题: Re: how to avoid contamination when doing PCR?
发信站: The unknown SPACE (Sun Sep 17 22:35:01 2000), 站内信件

biz is a master guy on pcr. i have a little to say. i have been using
tail DNA for pcr characterization of mice KO. my experience is that
u should separate all the reagents/pippets/pippetmen from commonly
used stuff. so, first of all, go to clean the pcr station, and make it
as clean as possible. remember, always NOT to bring template to the
station. also, get pcr buffer/ddH2O changed frequently if u question it.
primers should be stocked as 100uM, and u may only aliquot them for use.
use filtered pippet. change glove/pippet from time to time, the same as
u deal with RNA. there is a story in my lab. a guy tried RTPCR to clone
Drosophila PKR ( eIF2a kinase ), and got a product, which turned out
to be human PKR:). so u see what is happenning here.

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