A simplified method to prepare PCR template DNA for screening of transgenic and knockout mice

Polymerase chain reaction (PCR) amplification of DNA is the most widely use d technique for screening of large numbers of genetically engineered transg enic or knockout mice (Mus musculus). In this report, we present a new DNA preparation procedure for running diagnostic PCR, In this procedure, mouse ear tissue tvas used directly for PCR after the tissue underwent brief dige stion in a solution containing only proteinase Ii, Using this method, we ha ve successfully screened several Lines of single, double, and triple transg enic and knockout mice, The results are reliable and reproducible. The adva ntage of this new method is that DNA purification by organic extraction or isolation kit was omitted. DNA purification is the Limiting factor in terms of time and money when screening transgenic and knockout mice by PCR, In a ddition, using ear instead of tail tissue can reduce distress of animals be cause the samples can be obtained when the mice are labeled by ear punch.