Promoter ligation activated transcription amplification of...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12N 15/10 (2006.01) C12N 15/11 (2006.01)

Patent

CA 2062963

2062963 9118115 PCTABS00008 This invention discloses a scheme for producing nucleic acid end products that are functionally or exactly identical to the starting products, thereby resulting in exponential amplification of a desired nucleic acide sequence. Specifically, sequences are cycled between RNA and DNA forms using the following basic steps: (1) a T7 RNA polymerase promoter is ligated onto a single-stranded DNA template; (2) T7 RNA polymerase makes many copies of RNA: (3) a complementary DNA is made from the RNA by extension of a primer by reverse transcriptase; and (4) the RNA template is removed by ribonuclease H. This amplification method is useful for purposes such as genetic research and diagnostic assays.

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