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临床血清循环miRNAs定量技术的建立
引用本文:桂俊豪,田亚平,温新宇,邓心新,董振南,贾兴旺,高静,闰玮,田丽媛. 临床血清循环miRNAs定量技术的建立[J]. 压电与声光, 2010, 0(4)
作者姓名:桂俊豪  田亚平  温新宇  邓心新  董振南  贾兴旺  高静  闰玮  田丽媛
作者单位:解放军总医院生化科;
基金项目:国家科技部基金资助(2006FY230300);;国家科技支撑计划项目(2009BAI8605)~~
摘    要:目的建立可靠的临床血清(浆)循环miRNAs定量技术。方法常规收集血清(浆)标本,mirVana PARIS试剂盒法抽提血清(浆)总RNA,采用DNaseI消化总RNA提取液,以miRNAs特异性茎-环引物引导反转录,通过TaqMan实时荧光定量PCR对U6及靶miRNAs进行检测。结果10份新鲜血浆总RNA浓度介于3.5~35.4ng/μl之间。对常规收集的400μl临床血清标本中U6、miR-16、miR-224均能实现特异扩增及定量,相应的平均Ct值约为30、25及32。6份不同留置时间血清标本总RNA浓度分别10.24和4.46ng/μl,定量PCR结果显示其中相应miR-16和miR-224的丰度却相对稳定。结论血清(浆)总RNA抽提及循环miRNAs定量切实可行。

关 键 词:血清  循环miRNAs  实时荧光定量PCR  

Establishment of platform for quantifying circulating miRNAs in clinical serum samples
GUI Jun-hao,TIAN Ya-ping,WEN Xin-yu,DENG Xin-xin,DONG Zhen-nan,JIA Xing-wang,GAO Jing,RUN Wei,TIAN Li-yu,epartment of Clinical Biochemistry,Chinese PLA General Hospital,Beijing,China. Establishment of platform for quantifying circulating miRNAs in clinical serum samples[J]. Piezoelectrics & Acoustooptics, 2010, 0(4)
Authors:GUI Jun-hao  TIAN Ya-ping  WEN Xin-yu  DENG Xin-xin  DONG Zhen-nan  JIA Xing-wang  GAO Jing  RUN Wei  TIAN Li-yu  epartment of Clinical Biochemistry  Chinese PLA General Hospital  Beijing  China
Affiliation:GUI Jun-hao,TIAN Ya-ping,WEN Xin-yu,DENG Xin-xin,DONG Zhen-nan,JIA Xing-wang,GAO Jing,RUN Wei,TIAN Li-yu,epartment of Clinical Biochemistry,Chinese PLA General Hospital,Beijing 100853,China
Abstract:Objective To establish reliable platform for quantifying circulating miRNA in clinical serum or plasma samples. Methods Serum(plasma) samples were collected routinely, and total RNA was extracted using mirVana PARIS kit and recovered total RNA preparations were treated with DNase I. Mamm small nuclear U6 and target miRNAs in total RNA preparation were reverse transcribed by specific stem-loop primers and real-time fluorescence quantitative polymerase chain reaction(rt-fqPCR) was adopted to quantify U6 and t...
Keywords:Serum  Circulating miRNA  Real-time RT-qPCR  
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