《ABI_real-time_PCR原理及应用》.pdf

荧光PCR 原理及应用 周煜 ZHOU Yu Application Specialist cnmcbsupport@ ABI公司及其荧光PCR仪器 美国ABI 公司荧光定量PCR仪发展历史 1995－世界上第一台定量PCR仪7700型 1997－向医院用户推出5700型定量PCR仪 2000 －推出7900型384孔荧光定量PCR仪 }公认最高端定量PCR仪 2001 － 7900型96孔荧光定量PCR仪 2001 －推出7000型荧光定量PCR仪取得巨大成功 2003 －获得了荧光定量PCR仪专利－确立ABI在业界内的独特地位 2004 －第3代荧光定量PCR仪7300和7500型 2007 －第4代荧光定量PCR仪StepOne和StepOnePlus 3 ABI 公司荧光PCR仪器 4 ABI 公司荧光PCR仪器 Applied Biosystems 7900HT Fast Applied Biosystems 7500 Fast Real-Time PCR System Real-Time PCR System Applied Biosystems StepOne™ Real-Time PCR System Applied Biosystems 7300 Applied Biosystems 7500 Real-Time PCR System Real-Time PCR System 5 各种荧光PCR仪器 6 荧光PCR的化学 两种荧光化学原理 ● Fluorogenic 5’ Nuclease Assay – An established and proven chemistry for high specificity nucleic acid real-time quantitation – The introduction of TaqMan® MGB (Minor Groove Binder) probes has produced a turn-key homogeneous chemistry for the detection of Single Nucleotide Polymorphisms (SNPs) ● SYBR® Green 1 Double Stranded DNA Binding Dye Assay – Ideal for target identification (screening) assays – A lower-cost alternative when high specificity is not required 8 TaqMan探针 5’ 3’ 5’ 3’ 3’ 5’ Excitation RR R Q Q Q Excitation R Q 3’