长爪沙鼠不同组织基因组DNA的提取条件比较

Comparison of Genomic DNA Isolation Methods in Different Tissues of Mongolian gerbil

目的建立长爪沙鼠基因组DNA的提取方法并对其脑、肝、肾和肌肉组织中的基因丰度进行比较。方法采用酚-氯仿法对不同消化温度和不同蛋白酶K浓度处理的各种组织提取DNA,比较其OD260/OD280值、DNA和蛋白质含量。结果提取DNA时温度和酶浓度对不同组织的DNA和蛋白质含量的影响不同。肾DNA样品中的蛋白质含量在低酶浓度组明显低于高酶浓度组,低酶浓度组的肾和肝DNA样品中的蛋白质含量均明显低于高酶浓度组。消化温度对提取肌肉组织的DNA含量影响较大,而从肾和肝组织中提取的DNA含量在37℃和50℃消化组之间没有差异。肝组织中的基因丰度最高,而肌肉组织中的基因丰度最低。结论提取DNA时,消化温度和酶浓度对提取不同组织的DNA和蛋白质含量有不同程度的影响。不同组织的基因丰度差异较大。

Objective To extract genomic DNA from different tissues of Mongolian gerbil and to compare the abundance of genes. Methods Using the phenol-chloroform method, genomic DNA was extracted from gerbil kidney, liver and muscle digested at different temperature and concentration of Proteinase K. The values of OD260/OD280, DNA and protein were detected. Results The digesting temperature and concentration of Proteinase K had different effects on extraction of genomic DNA from kidney, liver and muscle. Digesting at 37 ℃ and 50 ℃ made no difference on concentration of DNA extracted from kidney and liver, but proteins in low enzyme group were significantly lower than those in high enzyme group. The concentrations of DNA and protein of liver samples in low enzyme group were significantly lower than those in high enzyme group. Temperature had significant effect on DNA and protein concentrations of muscle samples. The abundance of genes in liver was higher than that in kidney and muscle. Conclusion Proteinase K concentration have effects on concentrations of genomic DNA and protein samples. The abundance of genes is variant in different tissues.