| 摘要 |
A preferred version of the present invention relates to a method of matching the Tms of a fixed, first primer and degenerate, second primer. The first primer has a completely known sequence. Thus, the Tm of the first primer can be readily calculated. The second primer has a known sequence portion and an unknown sequence portion. The second primer is actually a mixture of primers with each primer within sharing the same known sequence and having a different unknown sequence portion. The present invention enables the matching of the two Tm of the first primer to the Tm of the degenerate, second primer. The close matching of the two Tms greatly improves the efficiency of a PCR reaction with two such primers. Another preferred version of the invention relates to finding degenerate, second primers that occur at frequencies that differ from the frequency than would be expected. These two preferred versions can be combined to provide a method that has a fixed, first primer and a degenerate, second primer that have Tms that are matched. This combination also supplies a degenerate, second primer that is either closer to or farther away from the fixed, first primer than would be expected based on a random distribution of template nucleotides.
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