酒精和冷冻处理对肌肉组织基因组DNA提取的影响

提取基因组DNA的质量对于后续的基因扩增、酶切以及测序等分子生物学研究具有关键的影响,对肌肉组织进行不同的酒精和冷冻处理会显著影响提取DNA的质量。经过-80℃超低温冷冻数日的黄鳝样品解冻,取肌肉组织放入75%酒精处理后提取的基因组DNA完整性差、降解厉害;经过-80℃超低温冷冻数日的黄鳝样品直接取肌肉组织提取的基因组DNA比较完整、降解程度低;活体采取的黄鳝新鲜肌肉组织样品经过75%酒精处理,在-80℃超低温冷冻数日然后提取的基因组DNA质量也较好,电泳条带明晰而锐利,弥散降解带较少。后两种处理方法对黄鳝肌肉组织中DNA酶活性降低显著,因此提取的肌肉组织基因组DNA质量较高,可以有力支持后续的酶切、PCR扩增等分子生物学实验。

Effects of Ethanol and Cryopreservation on the Extraction of Genomic DNA from Muscle Tissue

The quality of extracted genomic DNA exhibits key effects on such molecular biological experiments as gene amplification,restriction endonuclease digestion and genome sequencing.Although traditional methods are effectively used to isolate genomic DNA from muscle tissues,yet different processing of muscle tissues of swamp eel with ethanol and cryopreservation have significant effects on the quality of extracted genomic DNA.The muscle tissues of swamp eel cryopreserved at-80 ℃ for several days were thawed and processed with 75% ethanol,from which genomic DNA extracted with traditional methods was severely degraded.However,genomic DNA directly isolated from tissue of swamp eel cryopreserved at-80 ℃for several days was slightly degraded and comparatively integral.In other experiment,fresh muscle tissue from sample of swamp eel were immersed in 75% ethanol and cryopreserved at-80 ℃ for several days,from which genomic DNA isolated was also of higher quality with sharp while slightly dispersed bands in electrophoresis detection.Therefore,the latter two experiments decreased the activity of DNAase and the extracted DNA of higher quality could be used for such molecular biology experiments as enzyme digestion and PCR amplification.