Competitive RT-PCR: Preparation of Competitor RNA.
نویسنده
چکیده
INTRODUCTION The first step in competitive RT-PCR is the synthesis and purification of the synthetic competitor. This is an RNA molecule designed to be reverse-transcribed and PCR-amplified with the same efficiency as the endogenous transcript of interest. Once the competitor molecule has been prepared, as described in this protocol, competitive PCR can be carried out, as described in Comptetitive RT-PCR: Estimation of Copy Number.
منابع مشابه
Competitive RT-PCR: Estimation of Copy Number.
INTRODUCTION In this protocol, sample and competitor RNAs (previously synthesized as described in Preparation of Competitor RNA for Competitive RT-PCR) are reverse transcribed (separately) in a pilot experiment. A constant amount of sample RT product is then combined with a 2-logserial dilution of competitor RT product for PCR. This procedure provides an approximate copy number for the sample, ...
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عنوان ژورنال:
- CSH protocols
دوره 2006 1 شماره
صفحات -
تاریخ انتشار 2006