| 摘要 |
The present invention relates to a method for selectively amplifying a target nucleic acid from a biological sample containing a mixture of the target nucleic acid and non-target nucleic acid, wherein the target nucleic acid is a double stranded, bluntended nucleic acid fragment with 5' phosphorylated ends and the target nucleic acid is less than the size of the non-target nucleic acid, comprising the steps of: a) mixing said biological sample, a 5' adapter and a 3' adapter, wherein the 3' adapter is complementary to the 5' adapter at the 3' end, thereby generating a double-stranded adapter complex; b) introducing a ligase to the mixture of step a) and ligating the 5' adapter of the double-stranded adapter complexes to target nucleic acid and non-target nucleic acid, thereby generating ligated complexes; c) heating the products of step b) to release the 3' adapter; d) adding a polymerase to fill in the single-stranded 5' protruding ends of the 5' adapter ligated to target nucleic acid and to the non-target nucleic acid; and e) adding 5' adapters that hybridize to the products of step d) and amplifying under conditions that favor amplifying target nucleic acid nucleic acid, whereby target nucleic acid is selectively amplified. |
