| 摘要 |
The present invention relates to an improvement in a method for amplifying a target sequence of a target polynucleotide. The method comprises the step of forming extension products of an oligonucleotide primer at least along the target sequence or along an extended oligonucleotide primer. The extension products are copies of the target sequence. The improvement comprises forming the extension products in the presence of a second polynucleotide, to which the oligonucleotide primer hybridizes except for 1-10 nucleotides at the 3'-end of the oligonucleotide primer. Under the conditions chosen, the oligonucleotide primer is extended along the second polynucleotide in a controlled manner relative to extension of such primer along the target sequence, thus providing a positive control for the amplification reaction, which control may be qualitative or quantitative. Optionally, a modified oligonucleotide primer is included in the amplification reaction. The modified primer is substantially identical to the oligonucleotide primer with the exception of a chemical modification at its 3'-end that prevents degradation, under the reaction conditions, of the 1-10 nucleotides referred to above. The method finds particular application in the area of nucleic acid amplification. |
