DNA Polymerase (1-2 kb/min) Enzyme (with 2.5 mM dNTPs)

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Catalogue No: abx071005
Price: US$261.00
(Size: 500 U)
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DNA Polymerase (1-2 kb/min) Enzyme (with 2.5 mM dNTPs)

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Datasheet SDS
DNA Polymerase is purified from E. coli expressing a cloned DNA polymerase from Thermus aquaticus. The enzyme consists of a single polypeptide with a molecular weight of approximately 94 kDa. DNA Polymerase has 5′-3′ DNA polymerase activity and 5′-3′ exonuclease activity. The extension rate is about 1-2 kb/min. The enzyme can amplify genomic DNA fragments up to 4 kb.

Contents:

Component 500 U 3 kU 10 kU
DNA Polymerase500 U 6 × 500 U 4 × 2500 U
10X Buffer 1.2 ml 6 × 1.2 ml 20 × 1.2 ml
2.5 mM dNTPs 800 µl 6 × 800 µl 20 × 800 µl
6X DNA Loading Buffer 1 ml 2 × 1 ml 4 × 1 ml


Target DNA Polymerase
Tested Applications PCR
Host Bacteria
Conjugation Unconjugated
Purity > 99% (SDS-PAGE)
Quality Control Assayed for amplication efficiency to amplify the p53 gene from 10 ng of human genomic DNA.
Storage Store at -20 °C for up to 2 years. Avoid repeated freeze/thaw cycles.
Molecular Weight 94 kDa
Buffer DNA Polymerase: Contains 200 mM Tris-HCl (pH 8.3), 200 mM KCl, 100 mM (NH4)2SO4, 20 mM MgSO4
Buffer: 20 mM Tris-HCl (pH 8.0), 0.1 mM EDTA, 1 mM DTT, 100 mM KCl, 50% glycerol, stabilizers.
Biological Activity One unit of DNA Polymerase incorporates 10 nmol of deoxyribonucleotide into acid-precipitable material in 30 minutes at 74 °C.
Endotoxin Level Functional absence of double and single stranded endonuclease activity.
Concentration 5 U/µl
Availability Shipped within 10-20 working days.
Note This product is for research use only.

Not for human consumption, cosmetic, therapeutic or diagnostic use.
Directions for use Reaction Components:

Component Volume Final Concentration
Template Variable as required
Forward Primer (10 µM) 1 µl 0.2 µM
Reverse Primer (10 µM) 1 µl 0.2 µM
10X Buffer 5 µl 1X
2.5 mM dNTPs 4 µl 0.2 mM
DNA Polymerase 0.5-1 µl 2.5-5 U
Nuclease-Free Water Variable N/A
Total Volume 50 µl N/A

Thermal Cycling Conditions:

Number of Cycles Temperature Time per Cycle
1 cycle 94 °C 2-5 min
30-35 cycles 94 °C 30 seconds
50-60 °C 30 seconds
72 °C 1-2 kb/min
1 cycle 72 °C5-10 min

Notes:
  • PCR products using this enzyme are not suitable for polyacrylamide gel electrophoresis (PAGE).
  • A final concentration of 2 mM MgSO4 is sufficient for amplification of most targets. For some targets, a higher Mg2+ concentration may be required.
  • For optimal results, we recommend using a 100 mM MgSO4 stock solution to prepare a titration from 2 mM to 4 mM (final concentration) in 0.25 mM increments.
  • 0.5 µl (2.5 U) enzyme is sufficient per 50 µl reaction volume. For better amplification, up to 1 μl (5 U) enzyme can be used.
  • If precipitation is observed after thawing the Buffer, warm in a 37 °C water bath to dissolve the precipitate and mix before use.
Research Articles on DNA Polymerase


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