通过梅花鹿P21基因的检测比较相对和绝对荧光定量PCR
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- 英文论文题名：Compare relative and absolute quantity PCR through sika deer P21 gene detection
- 论文作者：王大涛 ; 郭倩倩 ; 朱宏伟 ; 刘振 ; 李春义
- 英文论文作者：Wang Datao ; Guo Qianqian ; Zhu Hongwei ; Liu Zhen ; Li Chunyi ; Institute of special Wild Economic Animals and Plants ; Chinese Academy of Agricultural Sciences
- 年：2016
- 作者机构：中国农业科学院特产研究所;
- 论文关键词：实时荧光定量PCR ; 相对定量 ; 绝对定量 ; P21
- 英文论文关键词：real time fluorescent quantitative PCR ; relative quantification ; absolute quantification ; P21
- 会议召开时间：2016-06-01
- 会议录名称：“十二五” 鹿科学研究进展
- 语种：中文
- 分类号：S825
- 学会代码：ZXMY
- 学会名称：中国畜牧业协会
- 页数：5
- 文件大小：537k
- 原文格式：O
摘要
实时荧光定量PCR是具有高度灵敏性和特异性的核酸分析技术,以SYBR Green染料法为基础的相对定量和绝对定量检测方法,由于操作简单、成本较低而倍受青睐。本文以梅花鹿P21基因为待检测基因,β-actin作为内参基因,对不同浓度稀释的模板进行相对实时荧光定量PCR检测,然后分别对P21基因和β-actin基因进行绝对定量检测。结果表明,绝对定量的结果比相对定量结果更准确可靠,但是在模板浓度较低时两种方法都会出现较大误差。
Real time fluorescent quantitative PCR is a highly sensitivity and specificity nucleic acid analysis technology,relative and absolute quantification base on SYBR Green dye is using widely because of simple operation and low cost.In this paper,we detected P21 gene of sika deer using β- actin as internal control gene.Sample was diluted to different concentration and P21 and β- actin genes were detected through absolute quantification separately.The result show that absolute quantification is more accurate and reliable compared with relative quantification.Both methods shown big deviations when the template concentration is too low.
Real time fluorescent quantitative PCR is a highly sensitivity and specificity nucleic acid analysis technology,relative and absolute quantification base on SYBR Green dye is using widely because of simple operation and low cost.In this paper,we detected P21 gene of sika deer using β- actin as internal control gene.Sample was diluted to different concentration and P21 and β- actin genes were detected through absolute quantification separately.The result show that absolute quantification is more accurate and reliable compared with relative quantification.Both methods shown big deviations when the template concentration is too low.
引文
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[9]朱振洪,李金辉,万海同.不同荧光定量per分析方法检测大鼠脑缺血再灌注损伤中casoas-3基因的表达研究[J].激光生物学报.2011.20(3):7.
[2]Baigent SJ,Petherbridge LJ,Howes K,Smith LP,Currie RJ,Nair VK.Absolute Quantitation of Marek's Disease Virus Genome Copy Number in Chicken Feather and Lymphocyte Samples Using Real-Time Per[J].Journal of virological methods,2005,123(1):53-64.
[3]赵焕英,包金凤.实时荧光定量per技术的原理及其应用研究进展[J].中国组织化学与细胞化学杂志,2007,16(4):6.
[4]Zimmermann B,Holzgreve W,Wenzel F,Hahn S.Novel Real-Time Quantitative Per Test for Trisomy 21[J].Clinical chemistry,2002,48(2):362-363.
[5]Tsuji N,Kamagata C,Furuya M,Kobayashi D,Yagihashi A,Morita T,Horita S,Watanabe N.Selection of an Internal Control Gene for Quantitation of Mrna in Colonic Tissues[J].Anticancer research,2002,22(6C):4 173-4 178.
[6]孙红梅,邢秀梅,丛波,李春义,杨福合.鹿茸干细胞体外培养技术的研究[J].经济动物学报,2007,11(1):4.
[7]Kuhne BS,Oschmann P.Quantitative Real-Time Rt-Per Using Hybridization Probes and Imported Standard Curves for Cytokine Gene Expression Analysis[J].BioTechniques,2002,33(5):1078,1080-1072,1 084 passim.
[8]李敏,汪洋,李银萍,李娟,陈学平.Taqman探针与sybr Green实时定量per法检测转基因植物外源基因拷贝数的差异分析[J].安徽农业大学学报,2012,39(4):3.
[9]朱振洪,李金辉,万海同.不同荧光定量per分析方法检测大鼠脑缺血再灌注损伤中casoas-3基因的表达研究[J].激光生物学报.2011.20(3):7.