| 摘要 |
The invention relates to a method for amplifying a target nucleic acid sequence tn-tC-tV-tC'-tn' comprising a first amplification using a first primer pair with sequence ma-K-pC and ma-K'-pC', and a subsequent second amplification using a second primer pair with sequence aL-aP-aK and aL'-aP'-aK', wherein pC is the same sequence as sequence element tC. pC and pC' are 8 to 40 nucleotides in length, K comprises a 3'-terminal sequence k1-k2-s, s is a mismatch sequences preventing PCR bias, ak is the same sequence as sequence element k1, aP-aK hybridize to a contiguous sequence on sequence element ma-K, k1 is 2 to 9 nucleotides in length, aL and aL' can be any sequence, and tV is a variable region within said target nucleic acid sequence. The invention further relates to collections of primer sets for use in the method of the invention. |
