New method of measuring CDNA by PCR and

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A combination of reverse transcription (RT) followed by the polymerase chain reaction is useful for analyzing low levels of mRNAs, but it sometimes does not yield quantitative information especially when the amount of the target mRNA is very small. Some researchers have described competitive polymerase chain reaction in which DNA fragments containing the same primer template sequences as the target compete for primer binding and amplification. Here, we developed a means of examining the ratio of the short to the long form of mouse prolactin receptor cDNAs, by means of "one-sided competitive polymerase chain reaction" . The procedure consisted of polymerase chain reaction using a primer common to both forms and primers specific to each of them. In addition, a means of measuring the level of cDNA encoding the extracellular domain of prolactin receptor was developed using competitive polymerase chain reaction, to estimate amount of prolactin receptor cDNA in cDNA samples. Now we try to develop a detection method for mRNA on the tissue section.