Method for transcriptional amplification of nucleic acids combining steps of different temperatures

摘要

A method of transcriptional amplification includes: (a) obtaining a mixture by combining (i) a biological sample comprising nucleic acids, (ii) amplification primers, (iii) amplification reagents including enzymes required for amplification, and (iv) at least one polyol capable of stabilizing the enzymes required for amplification; (b) denaturing the nucleic acids by heating the mixture at a temperature above 41° C.; and (c) transcriptionally amplifying at least one target nucleic acid at a temperature above 41° C. when present in the mixture.

主权项

1. A method of transcriptional amplification, comprising:
a) obtaining a mixture by combining (i) a biological sample comprising nucleic acids, (ii) amplification primers, (iii) amplification reagents including enzymes required for amplification, and (iv) at least one polyol capable of stabilizing the enzymes required for amplification; b) denaturing the nucleic acids by heating the mixture at a temperature of 46° C. or more; and c) transcriptionally amplifying at least one target nucleic acid at a temperature of 46° C. or more when present in the mixture; wherein the at least one polyol is selected from the group consisting of lactose, sorbitol, sucrose, trehalose, and mannitol.