Development of fluorescent quantitative RT-PCR method for detection of Hey1 in bone marrow mesenchymal stem cells

LIU Qiang; et al

doi:10.13406/j.cnki.cyxb.2010.06.010

您当前的位置：

首页 >

文章列表页 >

Development of fluorescent quantitative RT-PCR method for detection of Hey1 in bone marrow mesenchymal stem cells

更新时间：2024-01-17

- Development of fluorescent quantitative RT-PCR method for detection of Hey1 in bone marrow mesenchymal stem cells
- Journal of Chongqing Medical University Vol. 35, Issue 6, Pages: 897-899(2010)
- 作者机构：
  
  重庆医科大学医学检验系临床检验诊断学教育部重点实验室
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.06.010
  CLC： R329
- Published：2010，
扫描看全文
[1]刘强,翁亚光,唐敏,罗敏.骨髓间充质干细胞Hey1基因表达水平荧光定量RT-PCR方法的建立[J].重庆医科大学学报,2010,35(06):897-899.

LIU Qiang, et al. Development of fluorescent quantitative RT-PCR method for detection of Hey1 in bone marrow mesenchymal stem cells. [J]. Journal of Chongqing Medical University 35(6):897-899(2010)
[1]刘强,翁亚光,唐敏,罗敏.骨髓间充质干细胞Hey1基因表达水平荧光定量RT-PCR方法的建立[J].重庆医科大学学报,2010,35(06):897-899. DOI： 10.13406/j.cnki.cyxb.2010.06.010.

LIU Qiang, et al. Development of fluorescent quantitative RT-PCR method for detection of Hey1 in bone marrow mesenchymal stem cells. [J]. Journal of Chongqing Medical University 35(6):897-899(2010) DOI： 10.13406/j.cnki.cyxb.2010.06.010.

摘要

目的:建立骨髓间充质干细胞中Notch信号通路靶基因Hey1表达水平的荧光定量RT-PCR方法。方法:培养骨髓间充质干细胞(C

)

提取总RNA

验证内参基因β-actin和目的基因Hey1的扩增效率

优化PCR条件

分析扩增曲线、融解曲线

凝胶电泳验证定量PCR产物。结果:内参基因β-actin和目的基因Hey1扩增效率一致（扩增曲线斜率差

0.1）

提示可以使用比较Ct法对Hey1进行相对定量分析;扩增曲线、融解曲线及凝胶电泳结果提示PCR产物特异性好。结论:本研究所建立的用于Hey1基因表达分析的定量RT-PCR检测方法

准确可靠

可用于Bmp9和Notch信号通路的研究。

Abstract

Objective:To develop the method of detecting the level ofHey1 expression_the target gene of Notch signaling pathway in bone marrow mesenchymal stem cells

using real-time fluorescence quantitative RT-PCR.Methods:The bone marrow mesenchymal stem cells (C2C12) were cultured and the total RNA was extracted.And then the amplification efficiency was verified between the reference gene β-actin and the target gene Hey1.To optimize the PCR conditions and analyse amplification curves and melting curve before validation of quantitatve PCR products with gel electrophoresis.Results:The reference geneβ-actin and the target gene Hey1 have the same amplification efficiency(slope difference of the amplification curve<0.1)

suggesting that you can use the comparative Ct method for analysing relative expression of Hey1.Through analysing the amplification curve

melting curve and gel electrophoresis results we can get that the qPCR product was very specific.Conclusion:The real-time quantitative RT-PCR methods established for detecting the expression of Hey1 gene is accurate and reliable

which can be used for studying the relationship of Bmp9 and Notch signaling pathways.

关键词

骨髓间充质干细胞Hey1荧光定量RT-PCR

Keywords

bone marrow mesenchymal stem cellsHey1Fluorescent quantitative RT-PCR

references

Views

257

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effect of TNF-α on adipogenesis of SD rat bone marrow mesenchymal stem cells and the related molecular mechanism

Effect of titin gene on the procession of MSCs differentiating into cardiomyocytes in vitro

Effects of autologous bone marrow mesenchymal stem cells on the renal cell proliferation and apoptosis and renal function in a rabbit of renal ischemia-reperfusion injury

Distribution of transplanted bone mesenchymal stem cells in colon of ulcerative colitis rat

Endosomatic transference of transplanted bone marrow mesenchymal stem cells in rat of ulcerative colitis

Related Author

Xie Youhong

Li Lin

WANG Jinju

et al

吕铁伟

张小飞

李扬

朱静

Related Institution

Center of Digestion,Affiliated University Town Hospital of Chongqing Medical University

Department of Gastroenterology,The First Affiliated Hospital of Chongqing Medical University

重庆医科大学附属儿童医院儿科研究所

Department of Cardiology,the Children’s Hospital,Chongqing Medical University

贵州省习水县人民医院外一科

⁰