Rapid DNA extraction and PCR-sexing of mouse embryos

We have devised a PCR-based sexing method that is quick, simple, and highly reproducible. DNA is first extracted from embryonic mouse yolk sac via a 1 5 min, two-step incubation procedure utilizing PCR-compatible proteinase K buffer. Without any further manipulation the lysate is subjected to 30 cycl es of PCR, optimized to run in less than 1 hr. The reaction includes multip lexed primer pairs for Sry and Myog (myogenin) that generate a male specifi c band of 441 bp and an internal control band of 245 bp, respectively. This robust method is used routinely in our laboratory and gives rapid genotypi ng results with 98% reliability and 100% accuracy. Mol. Reprod. Dev. 60: 22 5-226, 2001. (C) 2001 Wiley-Liss, Inc.