Reproducibility in the Quantification of mRNA Levels by RT-PCR-ELISA and RT Competitive-PCR-ELISA
نویسندگان
چکیده
منابع مشابه
Reproducibility in the quantification of mRNA levels by RT-PCR-ELISA and RT competitive-PCR-ELISA.
The use of reverse transcription (RT) PCR for relative quantitation of gene transcripts relies on the reproducibility of the individual RT, PCR and product measurement steps. Semi-competitive RT-PCR (RT-cPCR) uses an internal competitor template in the PCR step to improve quantitation. We have surveyed the reproducibility of RT, PCR, RT-cPCR and measurement, amplifying the glyceraldehyde-3-phos...
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Because the down-regulation by progesterone of cystic fibrosis transmembrane conductance regulator (CFTR) expression could be a useful specific marker to define the state of implant receptivity in endometrium, a competitive reverse transcription-polymerase chain reaction (RT-PCR) was developed for quantifying the CFTR mRNA concentration in human endometrial samples. A competitor RNA was constru...
متن کاملModeling and analysis of competitive RT-PCR.
The present studies demonstrate a theoretical and practical framework for the accurate quantitation of gene expression in RNA extracted from microscopic tissue samples. The approaches are developed around competitive RT-PCR techniques. Assay performance has been examined and validated at both the RT and PCR steps. Our analysis of RT transcription efficiency for a number of native and competitor...
متن کاملCompetitive RT-PCR: Preparation of Competitor RNA.
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 Comptetit...
متن کاملCompetitive 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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ژورنال
عنوان ژورنال: BioTechniques
سال: 1998
ISSN: 0736-6205,1940-9818
DOI: 10.2144/98244rr02