›› 2010, Vol. 37 ›› Issue (1): 69-73.

• 生物技术 • Previous Articles     Next Articles

Development of a Real-time RT-PCR Assay for Quantificationof Rabbit GAPDH Gene

GAO B1,2, YANG Xiao-nong1,2, YU Xue-hui1,2, LUO Wei1,2, HUANG He1,2   

  1. (1.College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China;2.Key Laboratory of Veterinary Medicine in Universities in Sichuan Province, Chengdu 610041, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-01-20 Published:2010-01-20
  • Contact: YANG Xiao-nong

Abstract: According to the published GAPDH gene’s CDS sequence in GenBank, a pair of primers was designed and synthesized. Through optimization of react system and conditions, the method for detection of GAPDH gene of rabbit by SYBR Green Ⅰreal-time RT-PCR was established successfully. The results show that the lowest copy number for detection of GAPDH gene with this method is 32 copies/μL. and there is a good linear relationship in a wide range from 3.2×101 to 3.2×107 copies/μL (r=0.999). The coefficient of variation (CV) of 5 different concentration of positive plasmids is 1.67% to 4.73% and 2.66% to 8.74% in intra-assay and in inter-assay respectively. The method established in this paper has the advantages of rapidity,high sensibility, high throughput and good repeatability, which provides a methodological basis for quantitative analysis on function gene of rabbit and the expression of some disease gene in rabbit when GAPDH gene is taken as a reference gene.

Key words: rabbit; GAPDH gene; real-time RT-PCR; reference gene

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