Chen, F. and C.A. Suttle. 1995. Nested PCR with three highly degenerate primers for amplification and identification of DNA from related organisms. BioTechniques 18:609-611.

This paper reports that three highly degenerate primers can be used in nested PCR, and that the second amplification can be done directly on DNA fragments excised from low melting temperature agarose. The approach provides rapid confirmation that the correct targets have been amplified. It is useful when only one internal probe sequence (or gene-specific primer) is available to confirm the identity of a PCR product, and a degenerate primer sequence must be derived from an amino acid sequence. This can occur, for example, when DNA must be amplified from a group of related organisms.