Establishment and evaluation of SYBR Green Ⅰ real-time PCR for quantitatively detecting TMEM39A gene

doi:10.3969/j.issn.1004-1524.2018.11.20

›› 2018, Vol. 30 ›› Issue (11): 1958-1964.DOI: 10.3969/j.issn.1004-1524.2018.11.20

• Biosystems Engineering • Previous Articles Next Articles

Establishment and evaluation of SYBR Green Ⅰ real-time PCR for quantitatively detecting TMEM39A gene

LI Xiangrong¹, WANG Xinglong^1,2, LI Qian^1,2, MA Ruixian^1,2, ZHANG Haixia¹, LI Qiongyi^1,2, MA Zhongren¹, FENG Ruofei^1,*

1. Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China;
2. Life Science and Engineering College, Northwest Minzu University, Lanzhou 730030, China

Received:2018-06-07 Online:2018-11-25 Published:2018-12-21

Abstract

Abstract: In order to establish a rapid simple and reliable method for TMEM39A gene detection, a SYBR Green Ⅰ real-time PCR method was developed with a pair of primers designed according to the published gene sequence of TMEM39A in GenBank, and the reaction conditions were optimized. The results showed that the Ct value of the assay linearly related to the standard plasmid in the range of 3.937×10⁸-3.937×10³ copies·μL^-1 and the standard curve correlation coefficient R² was 0.999. It also had good specificity and sensitivity. The reproducibility was high, and the variation coefficients of intra-assay and inter-assay were less than 1%. This method could detect the content of TMEM39A gene in different cells, so its generality was good. In this study, a rapid and accurate method for quantification of TMEM39A gene by SYBR Green Ⅰ real-time PCR was successfully developed.

Key words: transmembrane protein 39A (TMEM39A), SYBR Green I real-time quantitative PCR, specificity, sensitivity

CLC Number:

LI Xiangrong, WANG Xinglong, LI Qian, MA Ruixian, ZHANG Haixia, LI Qiongyi, MA Zhongren, FENG Ruofei. Establishment and evaluation of SYBR Green Ⅰ real-time PCR for quantitatively detecting TMEM39A gene[J]. , 2018, 30(11): 1958-1964.

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Establishment and evaluation of SYBR Green Ⅰ real-time PCR for quantitatively detecting TMEM39A gene

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