发信人: netbuggie (中原一点虫), 信区: Biology
标 题: Re: question about QuantRTPCR
发信站: The unknown SPACE (Sat Mar 3 02:04:28 2001), 站内信件

yeap, man, it is like that you first have to make a DNA
competitor that can be amplified with the same primer
sets. however, the size of the product is different than
the RTPCR product, normally it is 30-100bp less than
that. so it is an internal deletion of the RTPCR product.
then you add different amount of the competitor to the
RT product, and do PCR. run the gel through a machine,
sort of like HPLC. there should be three major products,
the cDNA, the truncated cDNA, as well as a hybrid of
the cDNA and the truncated form. so as the name, it is
more precise to determine the relative mRNA level from
the samples, assuming that RT can work the same efficient
for all of them. it is a typical way other than northern.

【 在 bigband (翻山过河走九州) 的大作中提到: 】
: hi, Ah P,
: can you tell me the basic principles of Quantitative PCR,
: what is the utility of this technique?
: thanks!
: 【 在 netbuggie (中原一点虫) 的大作中提到: 】
: : a question on cloning of QuantRTPCR DNA competitor, if anybody
: : has done it b4. what is the basic strategies to make that
: : competitor? i am using PCR/clone to make an internal deletion,
: : but do not know why the PCR cannot work for the clone, though
: : it seems to be a deletion.


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