【创意版】PCR技术及其应用.pptVIP

Real-Time PCR (quantitative PCR) Detect fluorescence for each cycle .......... * Real-Time PCR (quantitative PCR) Quanlity control by melting curve .......... * Fluorescent reporter probe Real-Time PCR .......... * Real-Time PCR Pathogen Diagnosis microRNA Diagnosis heredopathia Diagnosis .......... * Iso-Thermal PCR Iso-Thermal Fast Sensitive LAMP / RPA .......... * Loop-mediated isothermal amplification (LAMP) .......... * Recombinase polymerase amplification Two primers Quantitativable Sensitive Long Primers .......... * Application of PCR .......... * Application of PCR Selective DNA isolation Amplification and Quantification PCR in diagnosis of diseases PCR in forensic science .......... * .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu .iuu PCR技术在医学中的应用 医学分子细胞生物学研究方法 .......... * What is PCR How to PCR Why do PCR .......... * Developed in 1983 by Kary Mullis polymerase chain reaction (PCR) 1993年的诺贝尔化学奖 .......... * Animation of PCR .......... * PCR Reagents Template DNA Primers dNTP DNA Polymerase Buffer Mg2+ .......... * Questions Annealling DNA polymerase v.s. Why primers are essential for DNA polymerase? .......... * Initialization step Denaturation step Annealing step Elongation step Final elongation Final hold PCR procedure stages 20~40 Cycles Exponential amplification Leveling off stage Plateau .......... * Evolution of machines for PCR 1 2 3 4 .......... * PCR Optimization Contamination Hairpins Polymerase errors Magnesium concentration Non-specific priming Primer dimers Deoxynucleotides GC-rich Template Proofreading DNA Polymerase Pfu, KOD … DMSO, betaine Redesign Primers Hot-Start Polymerase .......... * PCR Optimization: Primer Design Softwares Primer premier: e-PCR (NCBI) Oligo 7: Primer3: Command line style design tool Multi-Oligo design for probes or primers Single paired primers design for PCR Avoid non-specific amplification by blast .......... * Primers for amplification of human b-a