Trizol法提取鸡肝组织总RNA及miRNA的条件优化研究  

Optimization of Trizol Method for Extracting Total RNA and miRNA from Chicken Liver Tissue

作　　者：陈紫薇 张吉龙 庄武超 郑晓彤 陈建飞 CHEN Ziwei;ZHANG Jilong;ZHUANG Wuchao;ZHENG Xiaotong;CHEN Jianfei(College of Biotechnology,Jiangsu University of Science and Technology,Zhenjiang,Jiangsu 212100)

机构地区：[1]江苏科技大学生物技术学院,江苏镇江212100

出　　处：《中国家禽》2024年第9期175-181,共7页China Poultry

基　　金：江苏省基础研究计划自然科学基金项目(BK20220648)。

摘　　要：试验旨在优化总RNA及miRNA的Trizol法提取条件。试验利用Trizol法提取鸡肝脏组织总RNA,在RNA的沉降阶段,将样品置于室温、4℃、-20℃和-80℃条件下分别放置10 min、30 min和60 min,另外将样品在-80℃条件下放置3 h、6 h、12 h,分别探究不同沉降温度和不同沉降时间对总RNA完整度、纯度、质量浓度以及miRNA提取量的影响。结果显示:不同沉降温度和不同沉降时间均不影响总RNA的完整性;沉降时间能显著降低RNA的A260/A280比值(P<0.05),但仍在1.90~1.95范围内;不同沉降温度和不同沉降时间均显著影响总RNA和miRNA的提取量(P<0.05)。研究表明,-20℃放置30 min是总RNA提取的适宜条件,-80℃放置3 h为miRNA提取的适宜条件,-80℃放置10 min、4℃放置30 min和-20℃放置60 min的miR⁃NA提取效果良好,适合短时间提取miRNA的相关研究。The objective of this study was to optimize the extraction conditions for both total RNA and miRNA using the Trizol method.The total RNA was extracted from chicken liver tissue using the Trizol method.During the RNA sedimentation stage,the samples were subjected to various temperatures(-80℃,-20℃,4℃and room temperature)for different durations(10 min,30 min and 60 min),in addition to being placed at-80℃for 3 h,6 h and 12 h,to explore the effects of different sedimentation temperatures and sedimentation times on the RNA integrity,purity,mass concentration and the miRNA extraction concentration.The results showed that different temperatures and sedimentation times did not affect the integrity of total RNA.Sedimentation times had significant impact on the A260/A280 ratio of RNA(P<0.05),although it remained within the range of 1.90 to 1.95.More-over,different sedimentation temperatures and sedimentation times had significant effect on the extraction yield of total RNA and miRNA(P<0.05).The results revealed that the optimal conditions for total RNA extraction were achieved at-20℃for 30 min,while the optimal conditions for miRNA extraction were achieved at-80℃for 3 h.Notably,favorable miRNA extraction outcomes were observed when subjected to temperatures of-80℃for 10 min,4℃for 30 min and-20℃for 60 min were suitable for short-term miRNA extraction study.

关 键 词：鸡 肝组织 总RNA MIRNA TRIZOL法 RNA纯度 RNA浓度 

分 类 号：S831.2[农业科学—畜牧学]