HotStart DNA polymerase without dNTPs (250 units)

HotStart DNA polymerase inhibits PCR by using pyrophosphate (PPi) to trap Mg²⁺ ions essential for PCR, then breaking the PPi down by using heat resistant enzyme pyrophosphatase (PPase) as the temperature increases while processing the PCR reaction

※This product is shipped in dry ice.

$99.00

Cat. No.

E-3150-1

Product Overview

▶ Principles of HotStart DNA Polymerase

HotStart DNA Polymerase is a new concept of DNA polymerase that utilizes chemical interactions between pyrophosphate (PPi) and pyrophosphatase (PPase).
While divalent cation magnesium (Mg²⁺) plays an important role in PCR, the pyrophosphate binds with Mg²⁺ to form a complex that inhibits the PCR.
However, the Mg-PPi complex will decompose into Mg²⁺ and 2Pi by the thermostable pyrophosphatase (PPase) during the PCR reaction, allowing increased reactivity with the by-product of the digested PPi, 2Pi.

Features and Benefits

Sensitivity & specificity
Reduced generation of non-specific responses and primer-dimer formation during the mixing of PCR components by inhibiting DNA polymerase activity at the room temperature
Efficiency
Improved yield of target DNA through its high sensitivity and specificity
Reproducibility
Reproducible results with uniform quality products for each batch under the ISO 9001 quality system

Application

HotStart PCR, PCR with complex genomic templates/low copy templates/cDNA
Multiplex PCR
Primer extension
SNP typing
Real-time PCR using dsDNA Binding dye
Multiple primer pairs and amplification of low copy template DNA

Experimental data

Figure. Multiplex PCR comparison of genomic DNA using 6 sets of primers.

Lane M : 100 bp DNA Ladder (Bioneer, Cat. No. 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

Technical/Specs

▶ Specifications

5' to 3' exonuclease activity	No
3' to 5' exonuclease activity	No
3' – A overhang	Yes
Fragment size	Up to 12 kb

▶ Components

• 10X reaction buffer : Tris-HCl, KCl, Pyrophosphate, pH 9.0

• 1X dilution buffer : 50% glycerol containing 50 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, stabilizers, pH 8.2

• 10 mM dNTPs mix : 2.5 mM of each dNTP

• 20 mM MgCl₂

▶ Concentration

250 Units (5U/μl)

▶ Storage conditions

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

▶ Storage 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.

Literature/Support

◈ Manual

• Manual_HotStart DNA Polymerase

◈ MSDS

• MSDS_10 mM dNTPs

• MSDS_20 mM MgCl₂

• MSDS_10X Reaction buffer for HotStart DNA polymerase

• MSDS_Dilution buffer for HotStart DNA polymerase

• MSDS_HotStart DNA polymerase

◈ Quality Assurance

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

• ISO 9001 – certificate

Ordering Info

Cat. No.	Product Description	Option		Price
E-3150	HotStart DNA Polymerase	250 U	with dNTPs	$E-3150
E-3150-1		250 U	without dNTPs	$E-3150-1
E-3151		1000 U	with dNTPs	$E-3151