摘要
为建立检测鸡钙敏感受体的实时荧光定量PCR检测方法,通过对鸡钙敏感受体保守区的PCR扩增后,连入pMD18-T载体,制备阳性标准品,进行实时荧光定量PCR。通过对扩增曲线、熔解曲线等试验数据进行分析,得出该检测方法具有良好的稳定性和较高的精确度。本研究成功建立了检测鸡心肌组织钙敏感受体mRNA表达的荧光定量PCR方法,为定量研究鸡钙敏感受体mRNA表达及其与相关疾病的关系奠定基础。
This study was to develop a real-time fluorescent quantitative reverse transcription polymerase chain reaction （RT-PCR） assay using SYBR Green I dye for the detection of calcium-sensing receptor （CaSR） in chicken. The assay comprised of amplifying a cDNA template generated from total RNA extracted from heart issue of chicken and established a standard curve using in vitro transcribed CaSR RNA. Furthermore, melting curve analysis showed no primer dimers or non-specific products were produced. The SYBR Green I RT-PCR was specific, sensitive, and reproducible for the quantification of calcium-sensing receptor in chicken. The study successfully established real-time PCR method to detect CaSR mRNA expression in the myocardial tissue of chicken for the quantitative study of CaSR mRNA expression and its association with diseases in chicken.
出处
《中国家禽》
北大核心
2014年第3期10-13,共4页
China Poultry
基金
国家自然科学基金项目(30972162)
