牦牛乳RNA的完整性及其硬脂酰去饱和酶(SCD1)基因表达分析

Research on RNA integrity and related genes in yak milk

对牦牛乳进行不同的处理及加工后,提取牦牛乳中的总RNA进行质量浓度、纯度和完整性的检测,进行逆转录-聚合酶链反应(Reverse Transcription-Polymerase Chain Reaction,RT-PCR),为牦牛乳样品的采集与保存RNA提供依据,是一种较好的非侵染方法。结果显示:40,50,60,70,80,90℃加热30 min后牦牛乳中总RNA的质量浓度和纯度较新鲜牦牛乳有显著差异;2016年4~9月采集的牦牛乳-20℃保存至2017年4月时牦牛乳中总RNA的质量浓度和纯度较新鲜牦牛乳有显著差异;牦牛乳冻干粉与牦牛乳酪中总RNA的质量浓度和纯度较新鲜牦牛乳也有显著差异。三种不同的处理及加工下牦牛乳中总RNA在1%的琼脂糖凝胶电泳中均未跑出条带。但40,50,60,70℃加热牦牛乳30 min,以及2016年各月份采集的牦牛乳保存较长和牦牛乳冻干粉中SCD1基因的扩增条带均清晰整齐,适用于进行后续的分子生物学试验。

Total RNA concentration, purity and completeness as well as RT-PCR results in yak milk were examined after different process- ing to provide data for the collection and preservation protocols of yak milk samples which was an good no-invasive procedure. The results showed that total RNA concentration and purity in yak milk after 30min heating at different temperatures had significant difference compar- ing fresh yak milk. The concentration and purity of total RNA in the preserved yak milk were collected in different months in 2016 and also had significant difference comparing fresh yak milk. The same result occured to yak milk freeze-dried powder and yak cheese. There was no RNA strip in 1% agarose gel electrophoresis under three different treatments. However, the RT-PCR of SCD1 gene from yak milk of 30 man heating at 40℃,50 ℃,60 ℃ ,70℃, preserved yak milk of different months in 2016 and yak milk freeze-dried powder had vivid？ampli- fication bands, which were suitable for being used for further molecular biology experiments.