2.qRT-PCR-生物分析检测技术课件.ppt

第二节 实时荧光定量PCR SYBR Green I 定量原理 确定初始模板的浓度: 初始DNA量越多, 荧光达到某一值（域值）时所需要的循环数越少Log浓度与循环数呈线性关系：根据样品扩增达到域值的循环数就可计算出样品中所含的模板量 PRIMER AND PROBE DESIGN Primers and probes must be carefully designed because of the costs associated with producing probes with different dyes at 5 and 3 ends. PE/ABD Primer Express software, which is specifically designed to select the primers and probes. The required parameters for well-designed primers and probe have been well built into the program. These parameters include a Tm for the probe that is 10°C higher than the primers, primer Tm is between 58