Patent ID: 8632977
Filing Date: 2014-01-21
Classification: C12Q

Abstract:
1. A method for improving the annealing specificity of a primer in nucleic acid amplification, comprising: (a) preparing a 3′-end portion of the primer having a hybridizing nucleotide sequence substantially complementary to a site on a target nucleic acid to hybridize therewith; (b) forming at the 5′-end of the 3′-end portion a regulator portion comprising at least three contiguous universal bases to have the lowest T (c) forming at the 5′-end of the regulator portion a 5′-end portion consisting of a pre-selected arbitrary nucleotide sequence substantially not complementary to any site on the target nucleic acid, whereby the primer has three distinct portions and the annealing specificity of the primer is improved by the presence of the regulator portion and the 5′-end portion, and wherein the primer has a general formula of 5′-X