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  3. [TP5000] ExcelTaq™ Hot Start II DNA Polymerase (5 U/μl, 500 U)

[TP5000] ExcelTaq™ Hot Start II DNA Polymerase (5 U/μl, 500 U)

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Description

The ExcelTaq™ Hot Start II DNA Polymerase is a mixture of an aptamer-based inhibitor and a recombinant thermo-stable Taq DNA polymerase designed for preventing or minimizing non-specific DNA amplification in PCR reaction. The inactivation of polymerase is achieved by a reversible binding of the aptamer to the polymerase at temperatures below 45°C. The aptamer inhibitor releases polymerase during normal PCR cycling. The aptamer-based inhibition omits the time-consuming initial activation step required by chemically modified or antibody-based hot start polymerases.

The high specificity and sensitivity of ExcelTaq™ Hot Start II DNA Polymerase allows sensitive detection from limited amount of DNA templates, such as 1 pg of cDNA or 1 fg of plasmid DNA. With a high DNA synthesis rate and high thermo-stability, the ExcelTaq™ Hot Start II DNA Polymerase allows reactions to be set up at room temperature and is suitable for common and specialized PCR applications


Features

  • Aptamer-based hot start PCR

  • Reversible enzyme inactivation

  • Omits extra enzyme activation step

  • Convenient for room temperature PCR set-up

  • High yield and specificity of target amplicons

  • Wide range of amplicon length (up to 10 kb)

  • High sensitivity (as low as 1 fg of plasmid)


Applications 

  • High specificity PCR

  • Generation of PCR products for TA cloning

  • Routine PCR, multiplex PCR, colony PCR, and RT-PCR


Storage

-20°C for 24 months 

Odoo - Sample 1 for three columns

Hot Start

ExcelTaq™ Hot Start II shows absolute amplicon when pre-incubation at 37˚C and 42˚C. 

Reactions were subjected to a pre-incubation at 37˚C or 42˚C for 10 minutes before initial denaturation step. General Taq DNA polymerase generated primer dimers and failed to amplify target due to enzyme activated at 37˚C or 42˚C.


Odoo - Sample 1 for three columns

High specificity

ExcelTaq™ Hot Start II DNA Polymerase shows high specificity on amplifying target DNA. 

The optimal annealing temperature of GAPDH primer set is 58˚C. Improper annealing temperature set at 52˚C may force primer-dimer formation. ExcelTaq™ Hot Start II DNA Polymerase eliminated primer-dimer and increased amounts of desired product. Taq DNA polymerase reduced target amplification due to primer-dimer formation at improper annealing temperature of 52˚C. 


Odoo - Sample 2 for three columns

High sensitivity

ExcelTaq™ Hot Start II DNA Polymerase shows high sensitivity to amplify from low amount of templates.

Each set of PCR reactions contained either 1 ng, 10 pg, or 1 pg of HeLa cell cDNA as templates. ExcelTaq™ Hot Start II DNA Polymerase successfully amplified targets from lower amount of templates, in comparison with hot-start DNA polymerases from other suppliers and general Taq DNA polymerase. 

Odoo - Sample 3 for three columns

High sensitivity

ExcelTaq™ Hot Start II DNA Polymerase amplifies from plasmid template amounts as low as 1 fg.

 Contents

Component

Volume    

Hot start II DNA Polymerase (5 U/μl)

100 μl      

10X HS Buffer

1 ml x 2



Storage Buffer

50 mM Tris-HCl (pH 8.0), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, stabilizer, 50% (v/v) glycerol


10X HS buffer

200 mM Tris-HCl (pH 8.8 at 25°C), 100 mM KCl, 100 mM (NH4)2SO4, 20 mM MgCl2, 1% Triton X-100


Unit Definition

One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid-insoluble material in 30 minutes at 74°C. 


Storage

-20°C for 24 months 

 

Recommended PCR Condition

 

Template

1 – 150 ng*

Forward primer

0.1 – 0.5 µM

Reverse primer

0.1 – 0.5 µM

10X HS Buffer

5 µl

dNTPs

0.2 mM (each)

Hot Start II DNA Polymerase

0.25 µl (1.25U)

ddH2O

to 50 µl

Total volume

50 µl

*Optimal amount of DNA template depends on the source and quality of DNA.
The amount of purified plasmid templates can be even less than 1 pg.

 

 

Recommended PCR Program

Steps

Temp.

Time

Cycles

Template denature

94°C

2 min

1

Denature

94°C

30 sec

25-40

Annealing

50-68°C**

30 sec

Extension

72°C

30 sec/kb

Final extension

72°C

1 min

1

**Optimal PCR condition varies according to primers’ thermodynamic properties.

 

Odoo - Sample 1 for three columns

[RP1000] ExcelRT™ Reverse Transcriptase

  • High yield

  • Thermostable, up to 50°C, during first strand synthesis

  • High processivity, generating cDNA up to 8 kb

  • Reduced RNase H ribonuclease activity

Odoo - Sample 3 for three columns

[TP1000] ExcelTaq™ Taq DNA Polymerase

  • 5'→3' DNA polymerase activity

  • 5'→3' exonuclease activity

  • None detectable 3'→5' exonuclease (proofreading) activity

  • Generates PCR products with 3’-dA overhangs

  • Thermo-stable-half life is more than 40 min at 95°C 

Odoo - Sample 2 for three columns

[TQ1200] ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX) 

  • High Stability

  • Fast Hot Start

  • High Sensitivity

  • Low Background / High Specificity 

  • Suitable for Fast Program 

  • Smart Blue Contrast Dye 

Odoo - Sample 3 for three columns

[TP1200] ExcelTaq™ 5X PCR Master Dye Mix

  • 5’→3’ DNA polymerase activity

  • No detectable 3'→5' exonuclease (proofreading) activity

  • Generates PCR products with 3'-dA overhangs

  • High yield PCR 

  • High reproducibility

  • Reduced  pipetting errors

  • Includes tracking dye for direct loading after PCR