HotStart DNA Polymerase

Unique enzyme-mediated Hot Start DNA Polymerase

Bioneer’s HotStart DNA polymerase uses an exclusive enzyme-mediated Hot Start PCR method that, unlike most other HotStart PCR chemistries, completely releases all polymerase activity during the first denaturation step. Top DNA polymerase is completely inhibited by pyrophosphate at temperatures below 70°C. However, at temperatures above 70°C, a thermostable pyrophosphatase initiates pyrophosphate hydrolysis and activates the DNA polymerase. This prevents the formation of non-specific products and primer-dimers during the reaction set-up process and results in improved PCR specificity.

Features and Benefits

Fast:More than three times more processive than standard Taq DNA Polymerase
High performance:Amplifies fragments up to 12 kb
Value:No license fee to pay!

]]> Enzyme Properties

5' to 3' exonuclease activity: No
3' to 5' exonuclease activity: No
3’ – A Overhang: Yes
Fragment Size: Up to 12 kb

Application

Hot Start PCR, PCR with complex genomic templates/low copy templates/cDNA
Multiplex PCR
Primer extension
SNP typing
Real-Time PCR using SYBR Green dye
Multiple primer pairs and amplification of low copy template DNA

Reagents Supplied

10 x Reaction Buffer : Tris-HCl, KCl, Pyrophosphate, pH 9.0
1 x Dilution Buffer : 50% glycerol containing 50 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, stabilizers, pH 8.2
dNTP Mix : 2.5mM of each dNTP
20 mM MgCl₂

Concentration

5 U/μl

Storage Conditions

50% glycerol containing 20 mM Tris-HCl, 0.5 mM EDTA, 1 mM DTT, 100 mM KCl, stabilizers, pH 8.0

Store Temperature

-20°C

Unit Definition

One unit is defined at the amount of enzyme that will incorporate 10 nmole of dNTP into acid-insoluble material in 30 minutes at 72°C.

Figure 1. Multiplex PCR comparison of genomic DNA using 6 sets of primers and 2 different DNA Polymerases.

Lane M: 100 bp DNA Ladder (D-1030)
Lane 1: 750 bp fragment
Lane 2: 590 bp fragment
Lane 3: 450 bp fragment
Lane 4: 360 bp fragment
Lane 5: 260 bp fragment
Lane 6: 150 bp fragment
Lane 7: Multiplex PCR with primers used for Lane 1 – 6

]]> Manual

• HotStart DNA Polymerase

MSDS

• Hotstart DNA Polymerase

• Hotstart DP dilution buffer

• Hotstart DP reaction buffer

Brochure

• Enzymes 2010 Brochure

Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

• EC Directive 98/79/EC - certificate

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