摘要
建立了SYBR greenⅡ实时荧光定量PCR检测鲫鱼IgMmRNA水平的方法,即构建鲫鱼IgM标准品质粒,10倍稀释质粒标准品,建立标准曲线,进行熔解曲线分析和稳定性分析。结果表明,Ct值线性范围为5.8~25.4,相关系数为1;熔解曲线峰值单一,Tm为85.62(±0.13)℃;组内变异系数CV为0.18%~1.07%,组间变异系数为0.31%~2.31%。该方法具有检测范围广、扩增效率高、特异性好、重复性和稳定性良好等特点。
A SYBR Green Ⅱ based real-time PCR (Q-PCR) for quantitative analysis of Crucian carp (Carassius auratus)IgM mRNA was developed. To establish the standard curve, The positive recombinant plasmids served as a standard ,following with the analysis of melting curve and the stability test. The results showed that the linear range of Ct value was from 5.8 ~25.4 with a good correlation coefficient (r=1) and there was a single peak with a 85.62(±0.13)℃ Tm in melting curve. The coeffecients of variation (CV) were 0.18%~ 1.07% for the intra-assay test and there were 0.31%~2.31% in the inter-assay test.The real-time PCR assay for Crucian carp IgM mRNA has broad range, high efficiency, specificity and reliability.
出处
《广东农业科学》
CAS
CSCD
北大核心
2010年第6期185-187,202,共4页
Guangdong Agricultural Sciences
基金
农业科技成果转化资金项目(2008GB23260393)
现代农业产业技术体系建设专项(nycytx-49-14)