Top : Molecular Biology : PCR : Real-Time PCR

• Real-Time PCR Primer
• RT-PCR

Protocols

• Real-Time PCR (qPCR) Basics (Rob Powell, Brooke Laboratory, RCMB, Southampton General Hospital)
  A good place for you to learn real-time PCR. Topics included are as follows: Introduction to 'Real time' PCR Assay parameters Melt Curves and Primer Dimers Standard Curves Dynamic Range Quantification methods Introduction Delta CT Standard Curve
  http://www.primerdesign.co.uk/Download%20material/...
  Added: Sat Jan 31 2009, Hits: 58209, Reviews: 0  Write review  
• Real-Time PCR (M.Tevfik Dorak, MD, PhD)
  Excellent guide to real-time PCR including background; TaqMan primer and probe design guidelines; General recommendations for real-time RT-PCR;Setting up one-step TaqMan reaction;Cycling parameters for one-step PCR;Interpretation of results; The comparative CT method (DDCT) for relative quantitation of gene expression; Points to remember and trouble shooting; and more... (alternative url: http://home.att.net/~dorak/genetics/realtime.html)
  http://dorakmt.tripod.com/genetics/realtime.html
  Added: Sun Sep 22 2002, Hits: 40006, Reviews: 0  Write review   Cached
• SYBR Green Quantitative PCR Protocol (Lusis Lab, UCLA)
  Quantitative PCR is a method used to detect relative or absolute gene expression level. All qPCR involves the use of fluorescence to detect the threshold cycle (Ct) during PCR when the level of fluorescence gives signal over the background and is in the linear portion of the amplified curve. This Ct value is responsible for the accurate quantization of qPCR.
  http://www.genetics.ucla.edu/labs/lusis/greenquant...
  Added: Sat Mar 21 2009, Hits: 32609, Reviews: 0  Write review   Cached
• Essentials of Real-Time PCR (Life Technologies)
  Things you must know before you do qPCR including basics about Real-Time PCR Assays and Sequence Detection Chemistries SYBR� Green I Dye Chemistry About Quantitation Assays Absolute vs. Relative Quantitation Standard Curve Method for Relative Quantitation Comparative CT method for Relative Quantitation Standard Curve Method for Absolute Quantitation
  http://www.invitrogen.com/site/us/en/home/Products...
  Added: Thu May 02 2013, Hits: 4714, Reviews: 0  Write review   Cached
• Important Parameters of Quantitative PCR (qPCR) Analysis (Life Technologies)
  Very useful information about qPCR such as amplification efficiency, Setting the Baseline, Determination of Copy Number,Tm Calculations, PCR Optimization, etc.
  http://www.invitrogen.com/site/us/en/home/Referenc...
  Added: Thu May 02 2013, Hits: 3298, Reviews: 0  Write review   Cached
• Protocol for Real-Time RT-PCR (Dr. Xiaowei Wang at the Department of Molecular Biology, Massachusetts General Hospital)
  This protocol describes the detailed experimental procedure for real-time RT-PCR using SYBR Green I. The procedure begins with reverse transcription of total RNA. The cDNA is then used as template for real-time PCR with gene specific primers. You may need to modify this protocol if you use different reagents or instruments for real-time PCR.
  http://pga.mgh.harvard.edu/primerbank/PCR_protocol...
  Added: Fri May 14 2004, Hits: 2091, Reviews: 0  Write review   Cached
• Quantitative Reverse-Transcription PCR (q-RT-PCR)
  http://www.utexas.edu/research/sissonlab/protocols...
  Added: Sat Jan 31 2009, Hits: 17743, Reviews: 0  Write review  
• FAQs on Real-Time RT-PCR (Dr. Xiaowei Wang at the Department of Molecular Biology, Massachusetts General Hospital)
  Some questions and answers on real-time PCR primer design and amplification
  http://pga.mgh.harvard.edu/primerbank/faq.html
  Added: Fri May 14 2004, Hits: 1067, Reviews: 0  Write review   Cached
• How to Reduce Primer Dimers in a LightCycler PCR (Roche Molecular Biochemicals)
  Offers optimization strategies to overcome primer dimer for real-time PCR.
  http://www.wzw.tum.de/gene-quantification/roche-pr...
  Added: Sun Sep 22 2002, Hits: 6782, Reviews: 0  Write review  
• overviews on quantitative kinetic RT-PCR (Dr. Michael W. Pfaffl, Gene Quantification )
  This review discusses the technical aspects involved, contrasts conventional and kinetic RT-PCR methods for quantitating gene expression and compares the different kinetic RT-PCR systems. It illustrates the usefulness of these assays by demonstrating the significantly different levels of transcription between individuals of the housekeeping gene family, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH).
  http://www.wzw.tum.de/gene-quantification/adope-pd...
  Added: Sun Sep 22 2002, Hits: 7878, Reviews: 1 Read reviews  Write review  
• Quantification using real-time PCR technology (Dr. Michael W. Pfaffl, Gene Quantification )
  The introduction of real-time PCR technology has significantly improved and simplified the quantification of nucleic acids, and this technology has become an invaluable tool for many scientists working in different disciplines. Especially in the field of molecular diagnostics, real-time PCR-based assays have gained favour in the recent past. However, the wide use of real-time PCR methods has also highlighted some of the critical points and limitations of these assays.
  http://www.wzw.tum.de/gene-quantification/klein-20...
  Added: Sun Sep 22 2002, Hits: 14472, Reviews: 0  Write review  
• Real-Time or Kinetic PCR (University of Iowa DNA Facility)
  This page describes the following topics on real-time PCR:
  • Real-Time Chemistry
  • Instrumentation
  • Real-Time PCR Quantitation
  • Primer and Probe Design 
  • Thermal Cycling Parameters 
  • Sample Preparation
  • Data and Analysis
  • Supplemental Information
  
  http://dna-9.int-med.uiowa.edu/realtime.htm
  Added: Sun Sep 22 2002, Hits: 7272, Reviews: 0  Write review   Cached
• Real-Time PCR Handbook (RRC Core Genomics Facility, University of Illinois at Chicago)
  Covers real-time PCR theories, methods, primer design, etc
  http://www.uic.edu/depts/rrc/cgf/realtime/start.ht...
  Added: Thu Feb 05 2009, Hits: 15507, Reviews: 0  Write review   Cached
• Real-time PCR in virology (Dr. Michael W. Pfaffl, Gene Quantification )
  This paper describes the background, advantages and limitations of real-time PCR
  http://www.wzw.tum.de/gene-quantification/mackay-2...
  Added: Sun Sep 22 2002, Hits: 6202, Reviews: 0  Write review  
• Real-Time PCR Quantification Using Cloned Standards and Multiple Housekeeping Genes
  We reported a general laboratory procedure for QRT-PCR, which addresses most of the inherent difficulties with this technique, can be easily adopted by any laboratory.
  Added: Mon Feb 02 2009, Reviews: 0  Write review  
• Sensitive mRNA Quantification - Gene Quantification Page (Institute of Physiology)
  Main focus of the GENE QUANTIFICATION web page is, to describe and summarize all technical aspects involved in quantitative gene expression analysis using real-time (kinetic) PCR & RT-PCR. e.g. detection chemistry, assay specificity, sensitivity, reproducibility, robustness, intra- & inter-assay variations, kinetic PCR efficiency calculation, quantification strategies, optimization strategies, data aquisition, data calculation, data normalisation, and statistics. To obtain high fidelity, accuracy and reliability in reverse transcription and subsequent kinetic PCR highly defined reliable quantification strategies, calculation procedures and statistical models are needed.
  http://www.wzw.tum.de/gene-quantification/index.sh...
  Added: Fri Jan 16 2004, Hits: 4919, Reviews: 0  Write review  
• SYBR Green qPCR Protocol (Fero Lab, Fred Hutchinson Cancer Research Center)
  http://labs.fhcrc.org/fero/Protocols/Q-PCR.html
  Added: Sat Jan 31 2009, Hits: 13092, Reviews: 0  Write review   Cached