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PCR (Polymerase Chain Reaction) on ZAGENO

FFPE RNA Kit SKU: 10760050 bio-World
From $ 198.32 (50 U)
Sizes 2 (50 - 100 U)
Catalog IDs 10760050-1, 10760050-2
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Pfu DNA Polymerase MasterMix [2X] G-Biosciences
Templates Genomic DNA, Plasmid DNA, Viral DNA Sample Amount /
From $ 66.00 (40 Reactions)
Sizes 1 (40 Reactions)
Catalog IDs 786-817
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PrimeSTAR® GXL Premix TaKaRa
From $ 281.00 (200 Reactions)
Sizes 2 (200 - 800 Reactions)
Catalog IDs R051A, R051B
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2X One-Step RT-PCR Master Mix with Dual Dye Norgen Biotek
Templates Total RNA, mRNA Sample Amount /
From $ 243.00 (100 Reactions)
Sizes 3 (100 - 500 Reactions)
Catalog IDs 28242, 28273, ...
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Ribonuclease Inhibitor Sigma-Aldrich
Reagent Type Inhibitors Quantity 10000 Units, 2500 Units Applicable Processes In vitro transcription, Real-time PCR, cDNA synthesis
From $ 136.00 (2500 Units)
Sizes 2 (2500 - 10000 Units)
Catalog IDs R1158-2.5KU, R1158-10KU
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REDExtract-N-Amp™ Blood PCR Kit Sigma-Aldrich
Templates Genomic DNA Sample Amount /
From $ 33.50 (10 Reactions)
Sizes 4 (10 - 1000 Reactions)
Catalog IDs XNABS-1KT, XNAB-1KT, ...
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Pwo Master Roche Life Science
Templates Genomic DNA, Plasmid DNA Sample Amount /
From $ 221.00 (100 Reactions)
Sizes 1 (100 Reactions)
Catalog IDs 3789403001
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AMPIGENE® qPCR 1-Step Green Kit Hi-ROX Enzo Life Sciences
Templates Total RNA, mRNA
From $ 250.00 (200 Reactions)
Sizes 2 (200 - 1000 Reactions)
Catalog IDs ENZ-NUC109-0200, ENZ-NUC109-1000
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Expand™ High FidelityPLUS PCR System, dNTPack Roche Life Science
Templates Genomic DNA Sample Amount /
From $ 135.00 (125 Reactions)
Sizes 2 (125 - 500 Reactions)
Catalog IDs 4743725001, 4743733001
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SYBR® Green JumpStart™ Taq ReadyMix™ Sigma-Aldrich
Templates Genomic DNA, Plasmid DNA
From $ 44.50 (20 Reactions)
Sizes 3 (20 - 500 Reactions)
Catalog IDs S4438-20RXN, S4438-100RXN, ...
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Expand™ 20 kbPLUS PCR System Roche Life Science
Templates Genomic DNA Sample Amount /
From $ 190.00 (40 Reactions)
Sizes 1 (40 Reactions)
Catalog IDs 11811002001
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REDTaq® DNA Polymerase Sigma-Aldrich
Templates Genomic DNA Sample Amount /
From $ 44.69 (50 Units)
Sizes 3 (50 - 1000 Units)
Catalog IDs D4309-50UN, D4309-250UN, ...
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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