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PCR (Polymerase Chain Reaction) on ZAGENO

7500 Real Time PCR Systems Spectral Calibration Kit I Thermo Fisher Scientific
Reagent Type Calibration standard Applicable Processes Calibration Quantity 1 Kit
From $ 1,030.00 (1 Kit)
Sizes 1 (1 Kit)
Catalog IDs 4349180
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Tissue Ex-Amp PCR Kit (with Kodaq MasterMix) Applied Biological Materials
Templates Genomic DNA Fidelity Proofreading activity Amplicon Size /
From $ 85.00 (120 Reactions)
Sizes 1 (120 Reactions)
Catalog IDs G927
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ExCellenCT One-Step EvaGreen qRT-PCR Kit - iCycler Applied Biological Materials
Fluorescent Dye Included Templates / Amplicon Size < 150 bp
From $ 135.00 (100 Reactions)
Sizes 1 (100 Reactions)
Catalog IDs G917-iC
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ProbeMasterMix (2X) High ROX for qPCR Genaxxon Bioscience
Fluorescent Dye Not included Templates Genomic DNA, cDNA, Plasmid DNA Amplicon Size /
From $ 135.70 (2.5 ml)
Sizes 2 (2.5 - 12.5 ml)
Catalog IDs M3010.0100, M3010.0500
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REDTaq® DNA Polymerase Sigma-Aldrich
Templates Genomic DNA Fidelity No proofreading activity Amplicon Size /
From $ 46.30 (50 Units)
Sizes 3 (50 - 1000 Units)
Catalog IDs D4309-50UN, D4309-250UN, ...
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2X One-Step RT-PCR Master Mix Norgen Biotek
Templates Total RNA Fidelity No proofreading activity Amplicon Size < 1.5 kb
From $ 243.00 (100 Reactions)
Sizes 3 (100 - 500 Reactions)
Catalog IDs 28113, 28114, ...
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VetMAX™ NA and EU PRRSV Reagents Applied Biosystems
Fluorescent Dye Not included Templates Viral RNA Amplicon Size /
From $ 866.00 (100 Reactions)
Sizes 1 (100 Reactions)
Catalog IDs 4468465
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PCR Master Roche Life Science
Templates Genomic DNA, cDNA Fidelity No proofreading activity Amplicon Size < 2 kb
From $ 261.00 (200 Reactions)
Sizes 1 (200 Reactions)
Catalog IDs 11636103001
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MyTaq™ Blood-PCR Kit Bioline
Templates Genomic DNA Fidelity No proofreading activity Amplicon Size /
From $ 221.00 (250 Reactions)
Sizes 1 (250 Reactions)
Catalog IDs BIO-25054
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SD Polymerase Hotstart, 10 U/ul Bioron
Templates cDNA, Genomic DNA Fidelity No proofreading activity Amplicon Size > 20 – 30 kb
From $ 120.00 (200 Units)
Sizes 2 (200 - 1000 Units)
Catalog IDs 108902, 108910
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GreenHot Master Mix Bioron
Fluorescent Dye Included Templates cDNA Amplicon Size /
From $ 96.00 (200 Reactions)
Sizes 2 (200 - 1000 Reactions)
Catalog IDs 119205, 119225
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Phusion™ Flash High-Fidelity PCR Master Mix Thermo Scientific
Templates / Fidelity No proofreading activity Amplicon Size /
From $ 84.75 (100 Reactions)
Sizes 1 (100 Reactions)
Catalog IDs F548S
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PCR Amplification

The polymerase chain reaction (PCR) is an in vitro method used for the amplification of a specific DNA template to generate a large number of copies in the span of just a few hours. The development of the PCR method in 1983 by Kary Mullis replaced the old and slow cloning techniques which required days of work.

PCR Kits and Methods

Real-Time PCR

Real-time PCR is a technique that monitors DNA amplification in real-time rather than at the end of the PCR run, as is the case for End-Point PCR.

End-Point PCR

PCR enables amplification of specific DNA fragments with the help of a thermocycler and is a widespread molecular biology technique.

PCR Reagents

PCR requires five reagents; DNA/RNA template, DNA polymerase, primers (forward and reverse), dNTPs and PCR buffers.

The benefit of PCR

Another benefit of the PCR method is its high sensitivity; theoretically, a single copy of the template is enough to start the reaction. The sensitivity and speed make the PCR techniques useful in both basic research and for commercial applications like in vitro diagnostics, forensics, and quality control.

The basic principles of the PCR method

To start a PCR reaction the following is needed:

  • A template: a DNA or RNA strand that we want to amplify.

  • Nucleotides (dCTP, dGTP, dATP, dGTP): the building blocks from which DNA is made.Primers: short nucleotide strands that will anneal to the position that we want to start and end the amplification.

  • DNA polymerase: the enzyme catalyzing the reaction.

  • (Accessory additives like buffers and Mg2+)

Our cDNA synthesis Troubleshoot will guide you though any initial setbacks.

The amplification process:

  • Denaturation: the DNA molecule is split into two single strands of DNA by heating (carried out in a PCR thermocycler)

  • Hybridization: During a cool-down phase, the primers anneal to the target sequences of the single stranded DNA

  • Elongation: Polymerase extends the DNA molecule starting at the primer sequences, and so doubles the template numbers

  • The process is repeated from Step 1 achieving an exponential increase in the number of templates until the substrate runs out or the product accumulation is halting the reaction.

Check out our End-Point PCR Troubleshoot if you encounter any problems.

The past, present and future of PCR

PCR was originally developed to detect specific DNA sequences in a sample, and though this is still the most used application, many other applications have been developed over the past decades.
Nowadays there exist different types of PCRs e.g. End-Point PCR, qPCR (quantitative Real-time PCR), RT-PCR (Reverse Transcription PCR), Multiplex PCR, Nested PCR and Digital PCR. Depending on the polymerases used, the PCR is featured with special properties like a hot start mechanism or proofreading activity to enable the best results.

See our Real-Time PCR Troubleshoot as well as the other troubleshooting guides for tips to perfect your experiment.

Problems in the lab? Learn about methods and experiments in our Knowledge section or look up a specific Troubleshooting guide.

Video and Image Credits:

Video: DNA Learning Center/YouTube