Bioneer HotStart DNA Polymerase with dNTPs (1,000 Units)

Bioneer HotStart DNA Polymerase with dNTPs (1,000 Units)

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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.

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

Applications

• 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

Specifications

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.