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Molecular Biology / PCR / Variants of PCR / Real-time polymerase chain reaction

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In Molecular Biology, real-time polymerase chain reaction, also called quantitative real time polymerase chain reaction (qPCR) or kinetic polymerase chain reaction, is a laboratory technique based on polymerase chain reaction, which is used to amplify and simultaneously quantify a targeted DNA molecule. It enables both detection and quantification (as absolute number of copies or relative amount when normalized to DNA input or additional normalizing genes) of a specific sequence in a DNA sample.

The procedure follows the general principle of polymerase chain reaction; its key feature is that the amplified DNA is quantified as it accumulates in the reaction in real time after each amplification cycle. Two common methods of quantification are the use of fluorescent dyes that intercalate with double-stranded DNA, and modified DNA oligonucleotide probes that fluoresce when hybridized with a complementary DNA.

Frequently, real-time polymerase chain reaction is combined with reverse transcription polymerase chain reaction to quantify low abundance messenger RNA (mRNA), enabling a researcher to quantify relative gene expression at a particular time, or in a particular cell or tissue type.

Although real-time quantitative polymerase chain reaction is often marketed as RT-PCR, it should not be confused with reverse transcription polymerase chain reaction, also known as RT-PCR.

 Contents

1 Background

2 Real-time PCR using double-stranded DNA dyes

3 Fluorescent reporter probe method

4 Quantitation

5 Applications of real-time polymerase chain reaction

6 References

7 Further reading

8 External links

 

REAL TIME RT-PCR PROTOCOL 1. PURPOSE This protocol describes how ...
REAL TIME RT-PCR PROTOCOL. SOP #: M011. REVISION LEVEL: .1. PAGE: 2 of 5. 4.1.1 The workbench along with the tube racks and pipetmen should all be ...
pga.tigr.org/sop/RT-PCR.pdf

RT-PCR: The Basics

Advantages of real-time RT-PCR; methods, chemistries, ... SYBR Green is the most economical choice for real-time PCR product detection. ...
www.ambion.com/techlib/basics/rtpcr/index.html

Real Time PCR

Mar 24, 2008 ... Troubleshooting Real-time PCR Efficiency above 100%, Google, Page 1 ... real time pcr AND troubleshooting AND low fluorescence, Google ...
www.scribd.com/doc/2347651/Real-Time-PCR

Real-time quantitative RT-PCR (Taqman)
PCR protocol (multiplex reaction). Before the PCR can be performed on the .... e) Real-time quantitative PCR. Once each of the steps a-d have been carried ...
www.diabetesgenome.org/docs/Patti_Lab_Real-Time_qPCR_pr.pdf

Real Time PCR

Fairly recently, a new method of PCR quantification has been invented. This is called ¡°real-time PCR¡± because it allows the scientist to actually view the ...
www.bio.davidson.edu/Courses/Molbio/MolStudents/spring2003/Pierce/realtimepcr.htm

Real Time PCR Protocol

Protocol for Real-Time PCR ... You may need to modify this protocol if you use different reagents or instruments for real-time PCR. ...
pga.mgh.harvard.edu/primerbank/protocol.html

Protocol for Real-Time RT-PCR
Troubleshooting. Here I listed a few major causes for real-time PCR failures. Please read the PrimerBank Help page for more details. ...
pga.mgh.harvard.edu/primerbank/PCR_protocol.doc

Protocol for Real-Time RT-PCR
Analyze the real-time PCR result with the SDS 7000 software. Check to see if there is any bimodal dissociation curve or abnormal amplification plot.
http://pga.mgh.harvard.edu/primerbank/protocol.html

Quantitativereal-time PCR protocol for analysis of nuclear receptor signaling pathways.

Quantitative real-time PCR protocol for analysis of nuclear receptor signaling pathways. Angie L. Bookout and David J. Mangelsdorf [env] ...
www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1402222

GMO specific real-time PCR system
GMO specific real-time PCR system. Protocol for event-specific quantitation. of Bt11 in maize. Method development:. National Veterinary Institute (Norway) ...
gmo-crl.jrc.it/summaries/Bt11-protocol.pdf

Real-Time PCR [M.Tevfik DORAK]

See also qPCR tips and troubleshooting at qPCR Troubleshooting Guide by ABgene; Protocol Online qPCR troubleshooting; Ten Most CommonReal-Time PCR Pitfalls ...
dorakmt.tripod.com/genetics/realtime.html

QUANTITATIVE RT-PCR PROTOCOL (SYBR Green I)
Quantitative RT-PCR Protocol (SYBR Green I). 4. QUANTITATIVE REAL-TIME PCR (qRT-PCR). 1. Do qRT-PCR and test the selected primers ...
schnablelab.plantgenomics.iastate.edu/docs/resources/protocols/pdf/realtime_RT-PCR.2007.04.01.pdf

Bio-Rad iCycler & iQ Real-Time PCR Systems
iCycler iQ Real-time PCR Detection system. 2) Click on the Library module on the left panel. Select the View Protocol tab. Under Protocol Files, select ...
www.cosmobio.co.jp/support/technical/manual_SPA_20060904/manual/BioRadiCyclerSetup.pdf

Multiplex Real-Time PCR.

Multiplex Real-Time PCR. This protocol was adapted from "New Technology for Quantitative PCR," contributed by Wolfgang Kusser, Sandrine Javorschi, ...
www.cshprotocols.org/cgi/content/full/2006/1/pdb.prot4113

 

Real Time PCR Tutorial

There are several methods to quantitate alterations in mRNA levels using real time PCR. Let's look at the standard curve method first. ...
pathmicro.med.sc.edu/pcr/realtime-home.htm

Real-Time PCR Vs. Traditional PCR
Real-Time PCR Vs. Traditional PCR. Description. This tutorial will discuss the evolution of traditional PCR methods towards. the use of Real-Time chemistry ...
www.appliedbiosystems.com/support/tutorials/pdf/rtpcr_vs_tradpcr.pdf

Applied Biosystems Real-Time PCR Rapid Assay Development Guidelines
This tutorial will discuss recommended guidelines for designing and running. real-time PCR quantification and SNP Genotyping (Allelic Discrimination) ...
docs.appliedbiosystems.com/pebiodocs/04371093.pdf

Biocompare - Tools and Technologies for Real-Time PCR

Tools and Technologies for Real-Time PCR. Biocompare's qPCR Tutorial presents researchers with an overview of real-time qPCR, identifies the advantages and ...
www.biocompare.com/pcr/tutorial/qpcr/

Real-time PCR - A Tutorial by Joshua Gray, Ph.D.

Quantitative Real-time PCR, a Tutorial by Joshua Gray, Ph.D. - Assistant Professor, Rutgers University. PCR-based methods for the detection of changes in ...
joshuapgray.com/RealtimePCR/realindex.htm

Real-Time PCR

Real-time PCR is also the most commonly used method to monitor the ... In addition to this tutorial, our staff can help you with different aspects of your ...
qcom.etsu.edu/mbcf/pcr.htm

Real-Time PCR Tutorial

Quantitative Real-Time PCR, a Practical Tutorial by Joshua Gray, Pharmacology & Toxicology, Rutgers University, Piscataway, NJ. Introduction ...
www.rci.rutgers.edu/~gray/RealtimePCR/realmain.htm

Real-time PCR - OpenWetWare

Real-time PCR is a PCR technique used to quantify starting amounts of nucleic acid ... An excellent, detailed Q-PCR tutorial by Margaret and Richard Hunt, ...
openwetware.org/wiki/Real-time_PCR

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