Improved method for reverse transcription

Details Improved method for reverse transcription Improved method for reverse transcription

C - Chemistry, Metallurgy

12

P

C12P 19/34 (2006.01) C12N 15/10 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2211367

A method for preparing a cDNA from a mRNA using a reverse transcriptase wherein reverse transcription is performed at a temperature at which the mRNA does not take a secondary structure, for example, at a temperature of 45°C or more. The method is performed, for example, using a heat-labile reverse transcriptase in the presence of a substance exhibiting chaperone function having chaperone function such as saccharides. The method is performed, for example, in the presence of metal ions necessary for activation of the reverse transcriptase and a chelating agent for the metal ions such as a deoxynucleotide triphosphate. The method is capable of reverse transcription over the full length of mRNA template even if the mRNA is a long chain mRNA and, as a result, producing a full length cDNA.

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