藏獒基因组DNA提取及RAPD扩增体系的优化

Genomic DNA Extraction and Optimization of RAPD Reaction System of Tibetan Mastiff

采用改进的盐析法提取了藏獒血液基因组DNA,并对RAPD扩增体系进行了优化。结果表明,改进的盐析法所提取的藏獒基因组DNA完整、无降解,完全可以满足RAPD分析的需要;20μL的最佳扩增体系组成为2.50 mmol/L Mg2+、0.60 mmol/L dNTP、50 ng模板DNA、1.25U Taq酶和0.3μmol/L随机引物,最佳退火温度为36.5℃。

The method of improved salting out was used for whole blood genomic DNA extraction of Tibetan mastiff,and the RAPD reaction system of Tibetan mastiff was optimized.The results showed that the integrity and non-degradable genomic DNA of Tibetan mastiff could be gained by the method of improved salting out,and a total volume of 20 μL optimal RAPD reaction system of Tibetan mastiff consisted of 2.50 mmol/ L Mg2＋,0.60 mmol/ L dNTP,50 ng genomic DNA,1.25 U Taq DNA polymerase,and 0.3 μmol/ L arbitrary primers.The best renaturation temperature of RAPD analysis of Tibetan mastiff was 36.5℃.