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! TOC: Methods 1 Onno Bakker, Academic Medical Centre Amsterdam, NL Housekeeping Genes: A Gold Standard? 2 Weisser/ Schnittger, Klinikum Grosshadern München, Germany The choice of house keeping genes in MRD-quantification of t(8;21) positive AML 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 4 Jochen Wilhelm, University Giessen Estimation of Genome Sizes by Quantitative Real-Time PCR Applications Regulation and Development 5 N. Neubauer, University of Copenhagen, Biokemisk Afd., Copenhagen, Denmark Relative Quantification of Insulin Gene Expression on the LightCycler Using SYBR Green I 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 7 Jim Whelan, Plant Molecular Biology Group, University of Western Australia, Crawley, Australia Investigation of Mitochondrial Biogenesis in Plants using Quantitative Real-Time PCR 8 E. Veistinen, Turku University, Dept. Medical Microbiology, Kiinamyllynkatu 13, FIN 20520 Turku Quantification of Ikaros Family Isoforms by Real-Time PCR 9 P. Stordeur, Dep. Immunologie- Hematologie-Transfusion, Hopital Erasme, Brussels, Belgium Methods to quantify cytokine gene expression by Real-Time PCR Oncology 10 Dr. Bernard, Idahotech, Salt Lake City, USA quantitative profiling for breast cancer using DNA and RNA markers 11 Melanie Königshoff, University Giessen Quantification of HER-2/NEU Gene Copy Number in Breast Cancer Tissue 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 13 C. H. W. Klaasen, C. Wilhelmina Hospital, Dep. Of Med. Microbiology & Infectious Diseases Nijmegen, NL Relative Quantification of Human DNA in Feaces (stool) 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 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 16 Francisco Barros, INGO, Santiago de Compostela Gene Dosage Determination by Real Time PCR 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) 18 Karin Berg, Pathology, John Hopkins Medical Inst, Baltimore, USA Analysis of Bone Marrow Engraftment Following in Utero Bone Marrow Transplantation in a Can
