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Instruction manual anti-Taq high 2004

anti-Taq highTCP-101, 100 μL(Correspond to 500 U Taq DNA polymerase)
Store at -20 °C.

Contents[1] Introduction[2] Components[3] Protocol[4] Example[5] References

CAUTIONAll reagents in this kit are intended for research purposes. Do not use for diagnosis or clinical purposes. Please observe
general laboratory precautions and follow safety guidelines while using this kit.

JAPANTOYOBO CO., LTD.
Tel(81)-6-6348-3888
www.toyobo.co.jp/e/bio
tech_osaka@toyobo.jp

CHINATOYOBO (SHANGHAI) BIOTECH, CO., LTD.
Tel(86)-21-58794900.4140

FOR RESEARCH USE ONLY.

NOT FOR HUMAN OR DIAGNOSTIC USE.

www.toyobo.co.jp/e/bio[ 1 ] Introduction

Description
anti-Taq high is a highly purified neutralization monoclonal antibody to Taq DNA
polymerase. This product provides an antibody-mediated hot start PCR to enhance thespecificity and sensitivity of PCR. This antibody inhibits polymerase activity before theonset of thermal cycling, preventing primer dimer formation and non-specificamplification. At the first denaturation step of the thermal cycling, the antibody is quickly
inactivated and PCR proceeds. The antibody-mediated hot start method is significantly
more convenient to use than other hot start methods.
Taq DNA polymerase
Heating step > 70℃
Polymerase activity blocked

Anti-Taq antibody
Polymerase activity restored

Fig. 1. Principle of Blunting by KOD DNA polymerase

Features- Inhibits ≥95% polymerase activity at 45ºC
-No contamination of mouse genomic DNA examined by PCR-Enhances the specificity and sensitivity of PCR-Can be used when other hot start methods are difficult to perform (e.g. capillary PCR)-The polymerase can be reactivated quicker than with methods utilizing a chemicallymodified polymerase[ 2 ] Components

100 μL
1 mL

anti-Taq high*
10 x PCR Buffer***100 μL of anti-Taq high; corresponds to 500 U Taq DNA polymerase.
** <10 x PCR Buffer >10 mM Tris-HCl(pH8.3), 500 mM KCl, 15 mM MgCl2**PCR Buffers supplied with purchased Taq DNA polymerases can be used instead of
this buffer.

[3]Protocol

1.Preparation of polymerase-antibody mixture
Taq DNA polymerase*+ anti-Taq highor1U
0 . 2 μL

Room temperature, 5 min

Use as a polymerase solution**

Taq DNA polymerase* (5 U/μL)+ Equal volume of anti-Taq high*Taq-based high efficient PCR enzymes can be used.
**The polymerase-antibody mixture can be stored for long term at -20 °C.
JAPANTOYOBO CO., LTD.
Tel(81)-6-6348-3888
www.toyobo.co.jp/e/bio
tech_osaka@toyobo.jp

CHINA
TOYOBO (SHANGHAI) BIOTECH, CO., LTD.
Tel(86)-21-58794900.4140

FOR RESEARCH USE ONLY.

1NOT FOR HUMAN OR DIAGNOSTIC USE.

www.toyobo.co.jp/e/bio

2. Typical reaction set up
Component

Volume37.2-X μL

Distilled water
10 X PCR buffer

5 μL

2 mM dNTPs

5 μL

Forward primer (10 μM)

1 μL

Reverse primer (10 μM)

1 μL

Polymerase-antibody mixture (2.5U/μL)

0.8 μL

Template DNA

X μL

Total Volume
50 μL
Note: Pre-denaturation step is not necessary on PCR cycling.

[ 4 ] Example

Example 1. Application of the hot start PCR using the Taq based high efficient PCR
enzyme
The efficiency of anti-Taq antibodies were evaluated by the amplification of the human
β-globin gene (3.6 kb). The result indicates that anti-Taq high increases the specificity
and sensitivity of the PCR compared with the control reaction and PCR mediated hotstart using company A’s anti-Taq antibody.

1 2 3 M

M: λ/Hind III Marker
1: Taq-based high efficient PCR enzyme
2: Taq-based high efficient PCR enzyme + anti-Taq high
3: Taq-based high efficient PCR enzyme + anti-Taq antibody (company A)[ 5 ] References

1) R.T. D'Aquila, L.J. Bechtel, J.A. Videler, Eron JJ, P. Gorczyca, J.C.Kaplan,Maximizing sensitivity and specificity of PCR by pre-amplification heating. Nucleic
Acids Res. 19: 3749 (1991)
2) Q. Chou, M. Russell, D.E. Birch, J. Raymond, W. Bloch. Prevention of pre-PCR
mis-priming and primer dimerization improves low-copy-number amplifications. Nucleic
Acids Res. 20: 1717-23 (1992)
3) D.E. Kellogg , I. Rybalkin, S. Chen, N. Mukhamedova, T. Vlasik, P.D. Siebert, A.
Chenchik. TaqStart Antibody: "hot start" PCR facilitated by a neutralizing monoclonal
antibody directed against Taq DNA polymerase. Biotechniques. 16: 1134-7 (1994)

JAPANTOYOBO CO., LTD.
Tel(81)-6-6348-3888
www.toyobo.co.jp/e/bio
tech_osaka@toyobo.jp

CHINA
TOYOBO (SHANGHAI) BIOTECH, CO., LTD.
Tel(86)-21-58794900.4140

FOR RESEARCH USE ONLY.

2NOT FOR HUMAN OR DIAGNOSTIC USE.