Rapid Cycle Real-Time PCR -- Methods and Applications: Quantification

ISBN-13: 9783642623172 / Angielski / Miękka / 2012 / 223 str.

Carl Wittwer; Meinhard Hahn; Karen Kaul

Opis

Rapid cycle real-time PCR is a powerful technique for nucleic acid quantification and analysis that takes less than 30 minutes to complete. Fluorescence is automatically monitored each cycle and the amount of template quantified by advanced analytical methods, such as the second derivative maximum method. Immediately following rapid cycle PCR, melting curve analysis is performed to verify product purity with SYBR Green I and/or genotype with fluorescently-labeled hybridization probes (HybProbes or SimpleProbes). Rapid cycle real-time PCR is often cited as the most versatile, efficient method for nucleic acid quantification in research and climical studies. Molecular analysis has never been easier

Spis treści

Methods: Part 1: Onno Bakker, Academic Medical Centre Amsterdam, NL - Housekeeping Genes: A Gold Standard?- Part 2: Weisser/ Schnittger, Klinikum Grosshadern München, Germany - The choice of house keeping genes in MRD-quantification of t(8;21) positive AML.- Part 3: Ronald H. Lekanne-Deprez, Dep. of Anatomie & Embryologie, Amsterdam, The Netherlands - Quantification of mRNA Using Linear Regression of Log-Linear PCR Data-Points as an Alternative for the Standard Curve Approach.- Part 4: Jochen Wilhelm, University Giessen - Estimation of Genome Sizes by Quantitative Real-Time PCR; Applications; Regulation and Development.- Part 5: N. Neubauer, University of Copenhagen, Biokemisk Afd., Copenhagen, Denmark - Relative Quantification of Insulin Gene Expression on the LightCycler Using SYBR Green I.- Part 6: Jürgen Loeffler, Medizinische Klinik, Abt. II, Otfried-Müller-Str. 10, 72076 Tübingen, Germany - Quantification of T-Cell Receptor Excision Circle DANN Using Fluorescence Resonance Energy Transfer and the LightCycler System.- Part 7: Jim Whelan, Plant Molecular Biology Group, University of Western Australia, Crawley, Australia - Investigation of Mitochondrial Biogenesis in Plants using Quantitative Real-Time PCR.- Part 8: E. Veistinen, Turku University, Dept. Medical Microbiology, Kiinamyllynkatu 13, FIN 20520 Turku - Quantification of Ikaros Family Isoforms by Real-Time PCR.- Part 9: P. Stordeur, Dep. Immunologie-Hematologie-Transfusion, Hopital Erasme, Brussels, Belgium - Methods to quantify cytokine gene expression by Real-Time PCR; Oncology.- Part 10: Dr. Bernard, Idahotech, Salt Lake City, USA - Quantitative profiling for breast cancer using DNA and RNA markers.- Part 11: Melanie Königshoff, University Giessen - Quantification of HER-2/NEU Gene CopyNumber in Breast Cancer Tissue.- Part 12: Remedios Castelló Cros, Dpto. Bioquímica. Centro de Investigación, Hospital la Fe, Av. Campanar, 21, 46009 Valencia, Spain - Quantitative real-time reverse transcription-PCR assay for urokinase plasminogen activator, plasminogen activator inhibitor type 1, and tissue metalloproteinase inhibitor type 1 gene expressions in primary breast cancer.- Part 13: C. H. W. Klaasen, C. Wilhelmina Hospital, Dep. Of Med. Microbiology & Infectious Diseases Nijmegen, NL - Relative Quantification of Human DNA in Feaces (stool).- Part 14: Chung-Che (Jeff) Chang, Assistant Professor, Director, Hematopathology Fellowship and Molecular/Pharmacogenetics Lab., Dep. of Pathology, Medical College of Wisconsin, 9200 W. Wisconsin Ave., Milwaukee, WI 53226 - Real-time quatification of tumor load (t(14;18))in follicular lymphoma patients.- Part 15: P. Bolufer, Laboratorio de Biologia Molecular, Universitario La Fe, Valencia, Spain - Real time quantification of AML rearrangements (AML1/ETO and TEL/AML1 ) in the diagnosis and monitoring of acute leukemia; Genetics.- Part 16: Francisco Barros, INGO, Santiago de Compostela - Gene Dosage Determination by Real Time PCR.- Part 17: Elaine Lyon, ARUP Laboratories, Salt Lake City, USA - Deletions and duplications of the cytochrome p450 2D6 gene using a reference gene and competitor (Alison Millson).- Part 18: Karin Berg, Pathology, John Hopkins Medical Inst., Baltimore, USA - Analysis of Bone Marrow Engraftment Following in Utero Bone Marrow Transplantation in a Canine Model.- Part 19: Elaine Lyon, ARUP Laboratories, Salt Lake City, USA - Detection of trisomy 21 using SNP analysis and allele area quantification (Genevieve Pont-Kingdon); Microbiology.- Part 20: C. Drosten, Bernhard-Nocht-Institute of Tropical Medicine, Hamburg, Germany - Rapid Detection

Blurb

Rapid Cycle Real-Time PCR is a powerful technique for nucleic

acid quantification and analysis that takes less than 30 minutes

to complete. Fluorescence is automatically monitored

each cycle and the amount of template quantified by advanced

analytical methods, such as the second derivative maximum method.

Immediately following rapid cycle PCR, melting curve analysis is

performed to verify product purity with SYBR Green I and/or

genotype with fluorescently-labeled hybrdidization probes

(HybProbes or SimpleProbes). Rapid cycle real-time PCR is often

cited as the most versatile, efficient method for nucleic acid

quantification in research and climical studies. Molecular analysis

has never been easier!