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Quantification of human SUZ12 with SYBR Green Ⅰ real-time fluorescent polymerase chain reaction

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Author:
No author available
Journal Title:
WORLD CHINESE JOURNAL OF DIGESTOLOGY
Issue:
20
DOI:
10.3969/j.issn.1009-3079.2007.20.009
Key Word:
SUZ12;SYBR GreenⅠ;实时荧光定量PCR

Abstract´╝Ü 目的:建立SYBR GreenⅠ实时荧光PCR定量检测人类SUZ12基因的方法.方法:将SUZ12 RT-PCR扩增片段克隆入载体pGEM-T后,经测序鉴定正确后,进行纯化和定量及系列稀释,应用SYBR GreenⅠ实时荧光定量PCR检测SUZ12,建立标准曲线,熔解曲线分析产物的特异性.结果:该法检测的最低拷贝数为14.2,线性范围为1.42×101-1.42×108拷贝,相关系数r为-1.00,1.42×107拷贝/L标本的批内变异系数(CV)和日间CV分别为1.8%和2.8%.熔解曲线分析显示单一的峰,熔解温度(melting temperature,Tm)为(81.37±0.16)℃.结论:实时荧光定量PCR方法检测SUZ12基因,快速有效、灵敏度高、特异性好.

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