Optimization of DNA extraction method for genotyping in transgenic mice

Abstract:

Objective To explore the influence of optimized DNA extraction on genotyping of transgenic mice，and to compare it with commonly used reagent kits in the field of scientific research.Methods Based on our previous patent，we modified the DNA lysis buffer formula and experimental protocol. Genotyping and validation were carried out using musculin （a novel transcription factor）-transgenic mice and enhanced green fluorescent protein-transgenic mice.Results The DNA lysis solution was prepared immediately before use，and was composed of 100 μL 0.025 N NaOH，160 μL 0.5 M EDTA，and 40 mL ultrapure water. Each sample was mixed with 180 μL DNA lysis solution，at 100 ℃ for 30 min. Compared with the commonly used genotyping kits in relevant fields，the optimized DNA extraction method effectively shortened the genotyping time while ensuring the accuracy of identification，and moreover，the DNA lysis buffer preparation was simple and convenient，enabling more reaction times，and reducing reagent costs and workload considerably.Conclusion This study provides an economical，simple，and reliable DNA extraction technique for genotyping in transgenic mice，which deserves application and promotion.