Protocol and application of ethanol method to extract genomic DNA from transgenic mouse embryos

Abstract:

Objective:To optimize the method of genome extraction and to establish an effective,convenient and fast research platform which could identify and screen the transgenic mice on a large scale.Methods:The Genomic DNA was extracted by using the ethanol method.Results:The results of spectrophotometry and gel electrophoresis revealed that the concentration,purity and quality of the genomic DNA extracted through the ethanol method had no obvious difference compared with that extracted through the classical phenolchloroform method.The genomic DNA obtained by the two extraction protocols could both be digested completely and able to identify the knock-in mice by PCR amplification.Both the templates were well used in gene sequencing and Real-time PCR to detect the exogenous gene and the relative copy numbers. Conclusion:The DNA extracted through the ethanol method was superior in quality and had stable and reliable test results in transgenic mice identifying,digestion,gene sequencing and Real-time PCR amplification.