| 摘要 |
A method for increasing the number of polynucleotides containing sequences corresponding to a mRNA species present in a sample, comprises the steps of: (i) reverse transcription of the mRNA species using a heeled 5'-amplificatio n primer (FAP-RAND) and a heeled 3'-amplification primer (TAP-RT), wherein eac h primer sequence is unique, and either or each heel sequence includes a RNA polymerase promoter site, and the FAP includes a variable sequence, whereby the RNA is reverse-transcribed to produce double-stranded cDNA and then multiple cDNAs according to the variable sequence; and (ii) amplification of the cDNA using primers sufficiently complementary to the primers, i.e. FAP a nd TAP, within FAP-RAND and TAP-RT. In one embodiment, the method additionally comprises the step of: (iii) in vitro transcription, to produce RNA run-offs from either end of the amplicons.
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