PCR/Amplification

Hot Start PCR

Hot Start PCR techniques focus on the inhibition of polymerase activity during reaction preparation. By limiting polymerase activity prior to PCR cycling, non-specific amplification is reduced and target yield is increased. Common methods for Hot Start PCR include chemical modifications, wax-barrier methods, and inhibition by a taq-directed antibody.

Sigma's JumpStart Taq polymerase is an antibody inactivated hot start enzyme designed to minimize non-specific amplification while increasing target yield. Unlike other Hot Start methods (i.e. chemical inactivation), JumpStart Taq polymerase does not require a pre-incubation step prior to cycling because polymerase activity is fully restored during the first denaturation cycle of the PCR reaction.

Sigma JumpStart Taq Enzymes & ReadyMixes:

  • Greater Specificity & Increased Target Yield. JumpStart Taq antibody prevents non-specific amplification while increasing target yield.
  • Enhanced Sensitivity. JumpStart Taq DNA Polymerases provide superior amplification regardless of template concentration.
  • Flexibility to Meet Reaction Needs. Formulated for a variety of PCR needs including the ease of direct load with REDTaq reagents or the convenience of our ReadyMixes that only require the addition of primers, template, and water.
  • Convenience. Reduce set-up time, eliminate contamination concerns, and avoid long reactivation steps with JumpStart Reagents & Readymixes.
Product No. Product Name Application Format
Clear PCR Products (no gel-loading dye added)
D9307 JumpStart ™ Taq DNA Polymerase Universal PCR 2 tubes: Taq & buffer
D4184 JumpStart™ Taq DNA Polymerase Universal PCR 3 tubes: Taq, buffer, MgCl2
P2893 JumpStart™ Taq ReadyMix™ Universal PCR 1 tube: ReadyMix
D5809 JumpStart™ AccuTaq™ LA DNA Polymerase Long & Accurate PCR 2 tubes: Taq & buffer
Red PCR Products (gel-loading dye added to DNA Polymerase)
D8187 JumpStart™ REDTaq® DNA Polymerase Universal PCR 2 tubes: Taq & buffer
P0982 JumpStart™ REDTaq® ReadyMix™ Universal PCR 1 tube: ReadyMix
P1107 JumpStart™ REDTaq® (Lower Taq Concentration) Universal PCR 1 tube: ReadyMix
D1313 JumpStart™ REDAccuTaq® LA DNA Polymerase Long & Accurate PCR 2 tubes: Taq & buffer
Individual Reagents
A7721 JumpStart™ Taq Antibody Hot Start Reagent 2 tubes: Antibody & buffer
DNTPCA1 CleanAmp™ dNTP, 10 mM mix Hot Start Reagent 1 tube: Mix of 4 dNTPs
DNTPCA2 CleanAmp™ dNTP, 2 µmol each dNTP Hot Start Reagent 4 tubes: Each dNTP at 50 mM
DNTPCA10 CleanAmp™ dNTP, 10 µmol each dNTP Hot Start Reagent 4 tubes: Each dNTP at 50 mM

Technical Data


Market Leading Performance of JumpStart Taq Enzymes

Inactivated by a Taq-Directed Antibody, JumpStart Taq Provides Increased Sensitivity, Specificity, and Yield

 

Sensitivity, Specificity, Yield
Ten nanograms (even lanes) or 100 nanograms (odd lanes) total human genomic DNA was amplified with primers targeted to a 5kb section of b-globin. Each PCR was performed according to supplier's recommendations.

 

Greater Specificity & Increased Yield with JumpStart Taq

 

JumpStart Taq Delivers Greater Specificity JumpStart taq polymerase and other commercially available hot start enzymes were used to amplify a five kilobase fragment of b-globin. Taq enzymes were activated per supplier's recommendations.

 

Superior Amplification Over a Wide-Dynamic Range of Template

4 Nanograms
JumpStart REDTaq
Four nanograms of template DNA was amplified under standard PCR conditions (lanes 1-4) and Hot Start conditions using JumpStart REDTaq.
0.4 Nanograms
JumpStart REDTaq
Standard PCR conditions (lanes 1-4) and Hot Start conditions using JumpStart REDTaq were used to amplify 0.4 ng template DNA.

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JumpStart REDTaq - Direct Load & Hot Start PCR . . . A Winning Combination!

REDTaq DNA Polymerase is a unique blend of Sigma's high quality Taq DNA Polymerase combined with an inert red dye. The unique benefits of REDTaq DNA Polymerase include quick recognition and confirmation of appropriate mixing. Additionally, the red dye serves as a tracking dye and migrates at approximately the same rate as a 125 bp fragment.

Confirmation of Taq Polymerase Addition

REDTaq Recognition 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.

Loading Buffers or Tracking Dyes not Required

REDTaq Loading 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.

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Exceptional Performance with JumpStart REDTaq ReadyMix

Hot Start PCR in the Convenience of a ReadyMix

 

Superior Performance with the Convenience of a ReadyMix 200 ng Lambda phage DNA was amplified with Sigma's JumpStart REDTaq ReadyMix (odd numbered lanes) and Competitor I's Direct Load ReadyMix (even numbered lanes). Taq was activated per the supplier's recommendations.

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