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Product Description

Hot-Start Gene Taq is a heat resistant-DNA polymerase for hot-start PCR. Before entering the PCR cycle, the enzyme is activated by heat treatment at 95°C for 5 min.
This product is chemically modified “Gene Taq FP”, which is a modified Taq DNA polymerase in which the contamination of DNA derived from the host bacteria is controlled as much as possible. As with Gene Taq, Hot-Start Gene Taq does not have 5'-3' exonuclease activity. Also, having a terminal transferase activity, the PCR product obtained can be used for TA cloning.
This product contains two kinds of reaction buffers which are conventional 10 x Gene Taq Universal Buffer and 10 x Brilliant Buffer. The 10 x Brilliant Buffer has the effect of increasing the specificity of PCR and can be used only for the amplification of short strand DNA.

Features

・High specificity and high DNA yield.・Having a Hot-Start function; the reaction solution can be prepared at room temperature.・Most suited for multiplex PCR・High purity, with the contamination of host-derived DNA reduced as much as possible.

Applications

• Hot-Start PCR
• multiplex PCR

Molecular Weight	68 kDa
Concentration	2.5 units/µl
Storage Conditions	20 mmol/l Tris-HCl(pH 8.0), 100 mmol/l KCl, 0.1 mmol/l EDTA, 0.5% Tween 20, 1 mmol/l DTT, 50% Glycerol
Unit Definition	One unit is defined as the enzyme activity incorporating 10 nmoles of deoxynucleotide into acid-insoluble precipitates at 74°C for 30 min using activated calf thymus DNA as a primer/template.
Purity	No change was observed in the agarose gel electrophoresis pattern of DNA after reacting 10 units of this enzyme and 1 µg of λ/Hind III digest at 74°C for 1 hr.

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Product Components

Hot-Start Gene Taq (50 units)

Component	Quantity	Storage	Description
Hot-Start Gene Taq	50 units x 1	-20°C
dNTP Mixture (2.5 mmol/l each)	160 µl x 1	-20°C
10 x Gene Taq Universal Buffer (15 mmol/l Mg2+ )	1 ml x 1	-20°C
10 x Brilliant Buffer (20 mmol/l Mg2+ )	0.2 ml x 1	-20°C

Hot-Start Gene Taq (250 units)

Component	Quantity	Storage	Description
Hot-Start Gene Taq	250 units x 1	-20°C
dNTP Mixture (2.5 mmol/l each)	800 µl x 1	-20°C
10 x Gene Taq Universal Buffer (15 mmol/l Mg2+)	1 ml x 1	-20°C
10 x Brilliant Buffer (20 mmol/l Mg2+)	1 ml x 1	-20°C

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Usage

Example of use 1: Multiplex PCR

Multiplex PCR was performed using plasmid DNA as a template and nine pairs of primers. The two kinds of reaction buffers attached to the product were compared.

Lane M: OneSTEP Marker 11
Lane 1: Hot-Start Gene Taq Brilliant buffer
Lane 2: Hot-Start Gene Taq Universal buffer
Lane 3: N Corporation modified DNA polymerase

PCR reaction solution composition

Template DNA	2.5 µl
Primer Mixture	1 µl
Reaction Buffer	1X
dNTP Mixture	0.2 mM
Polymerase	0.3 U
total	50 µl

Thermal cycler conditions (ABI2720)

95°C	30 sec.	10 cycles
65°C	1 min.
72°C	1 min.
95°C	30 sec.	27 cycles
60°C	1 min.
72°C	1 min.
72°C	7 min.

Example of use 2: Shuttle PCR

Shuttle PCR was performed using HeLa DNA (50 ng) as a template and the three kinds of primers described below, for which amplification was difficult.

Primer (Strand length: 500 bp)

A-1	F	5'-AGTGGGCAGAGCCAGTCACT-3'
	R	5'-AGACTGGATGCCCAGCCTAA-3'
A-2	F	5'-CAGACTCTGGGGCTGCCCAT-3'
	R	5'-TCAAACACTGGCTTGGGGTA-3'
A-3	F	5'-CCTTGGAGCAATGAGCAATG-3'
	R	5'-CATCTTTCCAGGCTCCTTCC-3'

PCR reaction solution composition

Template DNA	50 ng
Primer (10 µM each)	1 µl
10 x Reaction Buffer	5 µl
dNTP Mixture	0.2 mM
Polymerase	0.25 U
Total	50 µl

Thermal cycler conditions (ABI2720)

95°C	5 min.
95°C	30 sec.	30 cycles
68°C	1.5 min.
68°C	3 min.
4°C

 

Results

When the Brilliant Buffer was used for Hot-Start Gene Taq, a highly specific amplification was achieved even by using low specific primers, and the target amplified product was obtained with a high yield.

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Data Sheets

SDS(Safety Data Sheet)

• Please go to the page in Japanese.

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Related Information

Note

• This product is for research use only.

Related Products

• Buffer

Comparison of Gene Taq series

Product	Characteristics	5' → 3' exonuclease activity	terminal transferase activity
Gene Taq	The amplification efficiency of DNA fragments not more than 1 kbp is especially high.	－	＋
Gene Taq NT	Gene Taq NT has the same functions as natural Taq DNA polymerase.	＋	＋
Gene Taq FP	Gene Taq FP obtained by further purifying Gene Taq to a higher degree of purity and by controlling the contamination by the host-derived DNA as much as possible. It is suitable for RAPD PCR which is affected by contamination by the genomic DNA.	－	＋

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