革胡子鲶MHC基因表达qPCR分析的引物设计与评估

Journal of Tianjin Agricultural University ›› 2016, Vol. 23 ›› Issue (4): 33-37.

• Researches and Scientific Notes • Previous Articles Next Articles

Design and Evaluation of Primers for Analysis on MHC Gene Expression Profiles of Clarias gariepinus by Using Real-Time Quantitative PCR Technique

LI Zhuan-zhuan¹, WANG Yan², GAO Jin-wei¹, WANG Xiao-mei¹, CHEN Cheng-xun¹, LI Tao³, MO Bao-lin¹

1. Tianjin Key Laboratory of Aqua-Ecology and Aquaculture, College of Fisheries, Tianjin Agricultural University, Tianjin 300384, China;
2. Tianjin Ocean Pal Carol Biotech CO, Ltd, Tianjin 300350, China;
3. College of Basic Science, Tianjin Agricultural University, Tianjin 300384, China

Received:2016-03-23 Online:2016-12-20 Published:2019-10-14

Abstract

Abstract: In order to analyze the MHC gene expression profiles in Clarias gariepinus by using real-time quantitative PCR（qPCR）technique, primer pairs for amplifying the MHC gene fragments of C. gariepinus were designed using Primer 5.0 software and then assessed by means of general PCR and qPCR amplification, respectively. The results of qPCR evaluation showed that melt curve of the primer pair named MHCF₁-MHCR₃ presented a single sharp peak, and standard curve displayed that its amplification efficiency was 99.3% and R² value was 0.998. The results indicated that the primer pair had good specificity and efficiency of amplification as well as met the criteria of qPCR amplification. This work laid a foundation for analyzing MHC gene expression of C. gariepinus by using qPCR technique.

Key words: Clarias gariepinus, major histocompatibility complex, real-time fluorescence quantitative PCR, melt peak, amplification efficiency

CLC Number:

S965.128

LI Zhuan-zhuan, WANG Yan, GAO Jin-wei, WANG Xiao-mei, CHEN Cheng-xun, LI Tao, MO Bao-lin. Design and Evaluation of Primers for Analysis on MHC Gene Expression Profiles of Clarias gariepinus by Using Real-Time Quantitative PCR Technique[J]. Journal of Tianjin Agricultural University, 2016, 23(4): 33-37.

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Design and Evaluation of Primers for Analysis on MHC Gene Expression Profiles of Clarias gariepinus by Using Real-Time Quantitative PCR Technique

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