Method for amplification of RNA
AssigneeShimadzu Corporation
InventorsHiroshi Tonoike, Naoyuki Nishimura
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Abstract
An object of the present invention is to establish a method for amplifying an RNA in a biologically-derived sample by preparing a reaction solution in which a nucleic acid synthesis is not inhibited even in the presence of various biologically-derived impurities and as well as to enable a convenient, rapid and highly sensitive analysis of the RNA in the sample. In a method of the present invention, a reaction solution having a pH higher than that of an ordinarily employed reaction solution and/or containing a polyamine and/or a sulfated polysaccharide.
Claims17
A one tube-two step polymerase chain reaction method for amplification of a target RNA in a biological sample comprising the steps of:
- a) providing in the tube a reaction solution for reverse transcription comprising a reverse transcriptase and at least one of a polyamine, a sulfated polysaccharide, a salt thereof, or a mixture thereof;
- b) incubating the biological sample under conditions that produce a reverse transcription product;
- and
- c) incubating the reverse transcription product with a subsequently added DNA synthesizing enzyme under conditions that produce a polymerase chain reaction product.
A one tube-one step method for amplification of a target RNA in a biological sample comprising the steps of:
- a) providing in the tube a reaction solution for a reverse transcription and a polymerase chain reaction comprising a reverse transcriptase, a DNA synthesizing enzyme and at least one of a polyamine, a sulfated polysaccharide, a salt thereof, or a mixture thereof;
- b) incubating the biological sample under conditions that produce a reverse transcription product;
- and
- c) incubating the reverse transcription product with the DNA synthesizing enzyme under conditions that produce a polymerase chain reaction product.
Dependencies are inferred from the claim text.