Deoxyribonucleic acid preparation in polymerase chain reaction genotyping of transgenic mice

Background and Purpose: In an attempt to find a rapid and reproducible meth od for routine polymerase chain reaction genotyping of transgenic mice, two novel approaches were developed. Methods: One approach allowed reproducible amplification from crude lysates of tail snips, using a thermally activated polymerase. In a second approac h, for situations in which non-invasive techniques are necessary, oral swab specimens were amplified after DNA extraction by use of an isolation kit. Samples from 10 transgenic factor VIII knockout mice were genotyped after p rocessing by use of these and other methods. Results: False-negative results were not obtained by use of the two novel a pproaches. Despite their relative simplicity, both approaches yielded resul ts comparable to those obtained by use of procedures known to be reliable, such as organic extraction and a DNA extraction kit. Conclusion: Both approaches are useful for PCR-amplification of DNA from ma mmalian sources.