Method for adjusting amplification efficiency of target polynucleotide

摘要

Disclosed is a method for adjusting the amplification efficiency of a target polynucleotide in the amplification of the target polynucleotide by PCR using primers (i) to (iii) below, the method comprising adjusting the amplification efficiency of the target polynucleotide by changing the quantity ratio of the primers (i) to (iii) below:
(i) a first primer which is able to be base-paired with the target polynucleotide;(ii) a second primer which is able to be base-paired with the target polynucleotide in competition with the first primer and from which extension reaction by PCR less occurs as compared to the first primer; and(iii) a third primer designed to allow for the amplification of the target polynucleotide in pairs with the first primer.

主权项

1. A method of adjusting the amplification efficiency of a target polynucleotide in the amplification of the target polynucleotide by PCR using primers (i) to (iii) below in the same amplification, the method comprising
adjusting the amplification efficiency of the target polynucleotide by changing the quantity ratio of the primers (i) to (iii) below: (i) a first primer which is able to be base-paired with the target polynucleotide; (ii) a second primer which is able to be base-paired with the target polynucleotide at the same region with which the first primer is configured to hybridize to and from which extension reaction by PCR occurs less as compared to the first primer; and (iii) a third primer designed to allow for the amplification of the target polynucleotide in pairs with the first primer, wherein the Tm value of the first primer is higher than the Tm value of the second primer, and the adjusting comprises changing the quantity of the third primer with respect to the total quantity of the first primer and the second primer.