Routine Amplification

Polymerase chain reaction (PCR) is a technique that results in exponential amplification of a target DNA sequence. Developed in 1983 by Kary B. Mullis, PCR has evolved into a fundamental research tool. By cycling through various temperatures, PCR results in the extension of a sequence–specific primer by a DNA polymerase such as DNA polymerase isolated from Thermus aquaticus (Taq). The DNA polymerase reads the template and incorporates a complimentary nucleotide yielding a newly assembled complimentary strand.

Routine PCR amplification is used to verify clone length, produce DNA for down stream applications and for experiments where fidelity and specificity are not a concern. Taq DNA polymerase can amplify clones up to 7 kb in length on average with yields in the range of 100 ng/µl. For more complex experiments, Taq DNA polymerase can be blended with enhancers or other enzymes for Long and Accurate PCR, Hot Start PCR or Quantitative PCR.

Sigma's broad product range of optimized enzymes and reagents for standard PCR include Taq DNA polymerase, buffers, ReadyMixes, and specialty blends. Discover the advantages today!

• Taq DNA Polymerase
• REDTaq DNA Polymerase
• Taq DNA Polymerase ReadyMixes
• Taq DNA Polymerase Kits

Taq DNA Polymerase is a specialized thermostable enzyme isolated from the thermophillic bacterium, Thermus aquaticus. The recombinant form of this enzyme is expressed in E. coli. This 94 kDa protein shows no detectable levels of contaminating endonucleases or exonucleases by SDS–PAGE. It has both 5' to 3' polymerase and exonuclease activity.

Taq is provided with buffer and either with MgCl₂ or packaged with a separate vial of MgCl₂ for ease of optimization. SuperPaks of DNA Polymerase and REDTaq DNA Polymerase include all reagents necessary for PCR except specific primers, template, and water. Each set includes Taq DNA polymerase at 5 units/µl or REDTaq DNA polymerase at 1 unit/µl, optimized 10X Reaction buffer available with or without MgCl₂, and an Ultrapure dNTP mix.

Product Name	Technical Bulletin	Product #
Taq DNA Polymerase with 10X Reaction Buffer		D1806
Taq DNA Polymerase with 10X Reaction Buffer without MgCl2 (separate vial of MgCl2 provided)		D4545
TaqSuperPak™ DNA Polymerase		D5938
TaqSuperPak™ DNA Polymerase without MgCl2		D5813

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REDTaq DNA Polymerase is a unique blend of Taq DNA Polymerase with an inert red dye. REDTaq DNA Polymerase provides the same great amplification as standard taq and does not interfere with downstream applications such as automated or manual sequencing and restriction digestion. Alternatively, the inert red dye can also be removed by standard purification methods.

• Same great performance as Taq DNA Polymerase in a more convenient format for high throughput applications
• Quick recognition and confirmation of appropriate mixing
• No loading buffers or tracking dyes necessary. Sample can be taken directly from reaction and loaded onto an agarose gel.

Product Name	Technical Bulletin	Product #
REDTaq DNA Polymerase		D4309
REDTaq Genomic DNA Polymerase		D8312
REDTaq DNA Polymerase SuperPak		D6063
REDTaq Genomic DNA Polymerase without MgCl2		D2812

Vial 1 contains no REDTaq, vial 2 has 2.5 units of REDTaq added to a 50 µl reaction volume, and vial 3 shows a REDTaq PCR reaction solution thoroughly mixed.

Samples may be added directly to an agarose gel after PCR without the addition of a loading buffer or tracking dye. The dye in REDTaq acts as a tracking dye migrating at approximately the same rate as a 125 bp fragment.

Comparison of yield for 1, 2, 3, and 7 kb DNA fragments using REDTaq (R) and standard Taq (T) DNA polymerases under identical PCR cycling conditions.

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Taq DNA Polymerase ReadyMixes contain Sigma's high quality Taq DNA polymerase, 99% pure dNTP's and buffer in a 2X optimized reaction concentrate. Reduce pipette steps and minimize the risk of contamination with our ReadyMix reagents; simply add template and primers. Sigma's Taq DNA Polymerase ReadyMixes are formulated to address various PCR needs and offer the benefits of direct load with REDTaq ReadyMix or packaged with a separate vial of MgCl₂ for ease of optimization.

Product Name	Technical Bulletin	Product #
ReadyMix Taq PCR Reaction Mix		P4600
ReadyMix Taq PCR Reaction Mix without MgCl2 (separate vial of MgCl2 provided)		P4475
REDTaq ReadyMix PCR Reaction Mix		R2523

Amplification of 1, 2, 3, and 7 kb fragments and a 4.5 kb human genomic DNA using ReadyMix Taq PCR Reaction Mix.

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Sigma's Taq DNA Polymerase Core Kits provide a complete enzyme system for routine amplification. The CORE-1 Kit contains all components necessary for routine amplification including 10X PCR buffer, 10X PCR buffer without MgCl₂, MgCl₂, PCR grade water, and dNTP mix. CORE-T kit contains the same components with the addtion of DNA Taq Polymerase. Discover the convenience of a kit specifically packaged to include all necessary PCR components and designed to address your PCR needs.

Product Name	Technical Bulletin	Product #
PCR Core Kit		CORE-1
PCR Core Kit with Taq DNA Polymerase		CORE-T

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Routine Amplification | Human Genomic DNA | Long & Accurate PCR Reagents | Hot Start PCR | RT-PCR | Quantitative PCR | Specialty Enzymes | Ultrapure Nucleotides | PCR Reaction Components | PCR Lab Equipment | DNA/RNA Molecular Weight Markers | Whole Genome Amplification