家兔H-FABP基因Real-time PCR方法的建立

Development of The Real-time PCR Approach for Detection of Rabbit H-FABP Gene

本研究根据Gen Bank上登录的家兔H-FABP基因CDS序列设计保守引物,建立了基于SYBRGreen Ⅰ染料技术的real-time PCR方法。结果表明:所构建的标准曲线回归方程为CtH-FABP=-3.151x+39.647,相关系数(r2)为0.999,批内、批间重复性测定的变异系数分别小于5.13%和7.64%,说明该方法具有快速、灵敏、特异性高、重复性好等优点。这一方法的建立为进一步研究H-FABP基因在不同品种家兔表达奠定了基础。

The conserved primer was designed according to the CDS sequence of Rabbit H-FABP gene avail?able in GenBank for this study, a SYBR Green I dye based on real-time PCR method is developed. The results show that the standard curve was CtH-FABP=-3.151x+39.647,the correlation coefficient (r2) was 0.999,The coeffi?cient of variation values for both intra-experimental and inter-experi-mental reproducibility ranged less than 5.13% and 7.64% respectively. The real-time PCR method established in the study has advantages of rapid, good in sensitivity, specificity and reproducibility, which will lay the foundation for further studying expression of H-FABP in different rabbit breeds.