高原地区大鼠耳蜗RNA提取条件优化  

Optimization of RNA extraction conditions from rat cochlea at high altitude

作　　者：师晶晶 巴桑[1] 普珍[1] 石艳[2] 安芳[3] 龚嘎蓝孜[1] SHI Jing-jing;BA Sang;PU Zhen(Tibet University Medical College,Lhasa 850000,China)

机构地区：[1]西藏大学医学院,西藏拉萨850000 [2]吉林大学药学院实验药理与毒理学教研室,吉林长春130021 [3]西藏自治区人民医院眼科,西藏拉萨850000

出　　处：《中国实验诊断学》2020年第2期283-286,共4页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金资助(818660531)(81860567);西藏自治区自然科学基金重点项目(XZ2019ZRG-26);西藏大学教学研究与改革项目(XZDXJXYJ201806);西藏大学医学院科研培育基金项目(2019XSZX01);西藏自治区高等教育教学改革研究重点项目(JG2019-07)。

摘　　要：目的探讨在高原地区采用AMBION抽提试剂盒提取RNA的最佳条件。方法采用AMBION抽提试剂盒,对126份大鼠耳蜗(每份耳蜗分为2组,实验组和对照组)提取RNA,实验组采用AMBION抽提试剂盒优化条件提取RNA,对照组按照AMBION抽提试剂盒说明书提取RNA。结果实验组和对照组均能成功提取大鼠耳蜗组织RNA,总RNA的提取量分别为:0.896±0.348μg,0.536±0.404μg(P>0.05);实验组RNA浓度高于对照组,分别为:0.147±0.047μg/μl,0.072±0.022μg/μl(P<0.05);实验组RNA纯度高于对照组,纯度指标A260/280分别为:2.096±0.044,1.900±0.113(P<0.05);实验组RNA完整性指标RIN值高于对照组,分别为:9.689±0.459,8.588±0.662(P<0.05);实验组和对照组28S/18S分别为:1.967±0.141,1.863±0.192(P>0.05)。结论在高原地区采用AMBION抽提试剂盒提取大鼠耳蜗组织RNA,需要优化提取条件,其RNA浓度、纯度和质量都能满足后续实验要求,且样本重复性好。Objective To explore the best conditions for extracting RNA using AMBION extraction kit in high altitude area.Methods AMBION extraction kit was used for extracting RNA from 126 rat cochlea(each cochlea was divided into 2 groups,the experimental group and the control group).The experimental group used AMBION extraction kit with the optimized condition extracting RNA.RNA was extracted from the control group according to the instructions of AMBION extraction kit.Results The RNA of rat cochlea tissue was successfully extracted by the experimental group and the control group,the total RNA extraction were 0.896±0.348μg and 0.536±0.404μg(P>0.05);The RNA concentration in the experimental group was higher than that in the control group:0.147±0.047μg/μl,0.072±0.022μg/μl(P<0.05);The RNA purity of the experimental group was higher than that of the control group,the purity index A260/280 were:2.096±0.044,1.900±0.113(P<0.05);The RNA integrity index RIN value of the experimental group was higher than that of the control group:9.689±0.459,8.588±0.662(P<0.05);28 S/18 Sbetween the experimental group and the control group were not statistically different:1.967±0.141,1.863±0.192(P>0.05).Conclusion The use of AMBION extraction kit to extract RNA from rat cochlea tissue in high altitude area requires optimized extraction conditions.The RNA concentration,purity,and quality can meet the requirements of subsequent experiments,and the sample repeatability is good.

关 键 词：高原 大鼠耳蜗 RNA提取 优化 

分 类 号：Q752[生物学—分子生物学]