Rapid Cycle Real-Time PCR -- Methods and Applications: Genetics and Oncology

ISBN-13: 9783642639654 / Angielski / Miękka / 2011 / 205 str.

W. Dietmaier; Carl Wittwer; N. Sivasubramanian

Opis

Rapid-Cycle Real-Time PCR is a powerful technique for nucleic acid amplification and analysis that often requires less than half an hour to perform. Samples are amplified by rapid-cycle PCR followed by immediate melting curve analysis in the same instrument. Melting curve analysis of PCR products with SYBR Green I often allows product identification without gel electrophoresis. Furthermore, in the presence of fluorescent hybridization probes, melting curves provide "dynamic dot blots" for fine sequence analysis, including single nucleotide polymorphisms (SNPs). The method is often cited as the most versatile, efficient method for nucleic acid analysis in research and diagnostics in the fields of genetics and oncology. Molecular diagnostics has never been easier

Spis treści

for Genetics and Oncology Volume Rapid Cycle Real-Time PCR Methods and Applications.- I Methods Useful in Genetics and Oncology.- Quantification of Cytokine mRNAs in Human Myocardial Biopsy Samples by Real-Time Quantitative PCR Technology Using the LightCycler Instrument.- Quantitative Two-Step RT-PCR for the Detection of Human ABCA1 Transporter on LightCycler Using Hybridization Probes and External Standards.- Quantification of Human Genomic DNA Using Retinoic X Receptor B Gene.- Genotyping by Guanosine-Dependent Quenching of Single-Labeled Fluorescein Probes.- Limitations of Melting Curve Analysis Using SYBR Green I — Fragment Differentiation and Mutation Detection in the CFTR-Gene.- SYBR Green I Analysis of the Trinucleotide Repeat Responsible for Huntington’s Disease.- II Applications in Genetics.- Parallel Genotyping of Different Genes: A Rapid Real-Time PCR Approach.- Detection of a Single Base Substitution in Single Cells by Melting Peak Analysis Using Dual-Color Hybridization Probes.- Rapid Screening for Five Major Cystic Fibrosis Mutations by Melting Peak Analysis Using Fluorogenic Hybridization Probes.- LightCycler PCR for the Polymorphisms —308 and —238 in the TNF AIpha Gene and for the TNFB1/B2 Polymorphism in the LT Alpha Gene.- Rapid Genotyping of 2-bp and 9-bp Deletion Mutations Using the LightCycler.- Genotyping of the Methionine-Valine Polymorphism at Codon 129 of the Human Prion Protein by Melting Point Analysis of Fluorescently Labeled Hybridization Probes.- Rapid Detection of Missense Mutations in the Prostatic Steroid 5?-Reductase Gene Using Real-Time Fluorescence PCR and Melting Curve Analysis.- III Applications in Oncolon.- Analysis of Microsatellite Instability by Melting Peak Analysis with BAT26 and BAT25 Specific Fluorescence Hybridization Probes.- Two Color Multiplexing and Typing of Human Papillomavirus Types 16,18 and 45 on LightCycler.- Quantitative Analysis of AML1-ETO Fusion Transcripts in t(8;21) Positive AML Using Real-Time RT-PCR.- Rapid Quantitative Detection of Free Cancer Cells in the Peritoneal Cavity of Gastric Cancer Patients with Real-Time CEA RT-PCR Using Hybridization Probes.- Quantitative Measurement of the mRNA Expression of the Tumor-Associated Antigen PRAME by Real-Time RT-PCR Using LightCycler and SYBR Green I Technology.- Expression Analysis of Telomerase-Genes hTERT and hTR by Quantitative PCR on LightCycler.- Measurement of MDR1 Gene Expression by Real-Time Quantitative RT-PCR Using the LightCycler Instrument.

Blurb

Rapid Cycle Real-Time PCR is a powerful technique for nucleic acid amplification and analysis that often requires less than half an hour to perform. Samples are amplified by rapid-cycle PCR followed by immediate melting curve analysis in the same instrument. Melting curve analysis of PCR products with SYBR Green I allow product identification without gel electrophoresis. Furthermore, in the presence of fluorescent hybridization probes, melting curves provide "dynamic dot blots" for fine sequence analysis, including single nucleotide polymorphisms. The method is often cited as the most versatile, efficient method for nucleic acid analysis in research and analysis in the fields of Genetics and Oncology. Molecular analysis has never been easier!