| 摘要 |
<p>An annealing control primer and uses thereof are provided, which annealing control primer(ACP) comprises at least one universal base or non-discriminatory base analog which is positioned between 3-terminal and 5-terminal portions, limits the primer annealing to the 3-terminal portion at a specific temperature, and inhibits annealing at the 5-terminal portion, so that the annealing specificity can be improved. The annealing control primer for improving annealing specificity in nucleic acid amplification comprises: (a) a 3'-terminal portion having a hybridizing nucleotide sequence substantially complementary to a site on a template nucleic acid to be hybridized therewith; (b) a 5'-terminal portion having the pre-selected arbitrary nucleotide sequence; and (c) a regulator portion which is positioned between the 3'-terminal and 5'-terminal portions, comprises at least one universal base or non-discriminatory base analog, and regulates an annealing portion of the primer dependent on the annealing temperature, wherein the pre-selected arbitrary nucleotide sequence in the 5'-terminal portion is substantially not complementary to any site on the template nucleic acid; the template nucleic acid is gDNA, cDNA or mRNA; the gDNA or cDNA is single or double-stranded DNA; the nucleic acid amplification is performed under a first and a second annealing temperatures; the first annealing temperature is 30 to 68 deg. C; and the second annealing temperature is 50 to 72 deg. C.</p> |
