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PCR

Polymerase Chain Reaction

In our opinion the most important methodological invention in molecular biology has been the polymerase chain reaction (PCR) as it has allowed a literal explosion of molecular biology and genetics advancements through the cloning of genes.

PCR Articles

Analysis of PCR Reactions

Factors for Successful PCR

PCR Based Site Directed Mutagenesis

PCR Contamination

PCR Enzymes

Primer Extension

PCR History

PCR Primer Design

PCR No Bands or Product

PCR Troubleshooting

Long Non-Specific PCR Products

Site Directed Mutagenesis

PCR Samples Template

Real Time PCR

Real Time PCR History

PCR Forum Topics

Primer dimer Hi, When doing normal PCR, I always get a strong band at the bottom of the gel ~50bp instead of the expected band ~2kb. I think I'm having a primer...
Asap I'm doing 2tube ARMS PCR. When I use both pairs of my primers(control&specific;), i can't see my product. but when i use only the specific primers i...
loading dye Has anybody ever come across a loading dye that ran at a different molecular size on an agarose gel than expected? Why is this happening?
Cresol Red/Sucrose Hi. Does anyone have experience using cresol red/sucrose as loading dye added to the reaction BEFORE PCR? Does cresol red/sucrose have to have a...
Amplification won't work! I'm trying to amplify a ~200bp segment from a ~4kb plasmid. A postdoc in my lab previously amplified the segment and gave me a stock of gel-extracted...
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We really recommend you read our excellent review of the History of the Polymerase Chain Reaction (PCR)

PCR is a primer-mediated enzymatic amplification of specifically cloned or genomic DNA sequences.  It has become a routine procedure in every molecular biology lab for manipulating and identifying genetic material.  New applications have now been generated with remarkable regularity.  These applications are transforming molecular biology.
In the post genomic era, PCR had become the method of choice to clone existing genes and generate a wide array of new genes by mutagenesis and recombination within the genes of interest.  The quick and easy availability of these genes is crucial for the study of functional genomics, protein structure function relationships, protein engineering, protein protein interactions, gene expression and essentially molecular evolution.

 

Diagram 1 - PCR Cycle Steps

PCR

Steps in a PCR Reaction

There are 3 steps in one PCR cycle reaction (see diagram 1 - PCR Cycle Steps). In step 1, the DNA is denatured at high temperatures (from 90 - 97 degrees celcius). In step 2, primers anneal to the DNA template strands to prime extension. In step 3, extension occurs at the end of the annealed primers to create a complementary copy strand of DNA. This effectively doubles the DNA quantity through the 3 steps in the PCR cycle. This cycle can repeat indefinitely theoretically, however normally constitutes 25-40 cycles.

PCR Guidelines:

For a successful PCR you need the following (read the following articles to improve and learn PCR):

Template DNA or sample starting material

PCR primers

PCR Enzymes

Factors for a Successful PCR

Post-PCR Analysis:

Troubleshooting PCRs

Analysis of PCR Reactions

 

Please read our review of the History of PCR

Articles and Information on PCR

PCR Polymerases

Avoiding PCR Contamination

PCR Primer Design Guidelines and Software

Review on the History and Development of the Polymerase Chain Reaction (PCR)

Also visit PCR Station

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Polymerase Chain Reaction - PCR

Other Types of PCR - Related:

Real-Time PCR

 

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