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Kelnow Fragment
More Information
CAT #
EBT-1021
QTY
200 unit (5 unit/㎕)
PRICE (W)
60,000
MANUAL
MSDS
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Description

 

DNA Polymerase I Large (Klenow) Fragment is a proteolytic product of E.coli DNA Polymerase.
Klenow Fragment lacks the 5' to 3´ exonuclease activity, but retains its 5' to 3´ polymerase, 3´' to 5´ exonuclease and strand displacement activities. The 5' to 3´ polymerase activity of Klenow Fragment can be used to fill in 5´-protruding ends with unlabeled or labeled dNTPs, to sequence single- or double-stranded DNA templates, for in vitro mutagenesis using synthetic oligonucleotides, for cDNA second-strand synthesis and to generate single-stranded DNA probes. The 3' to 5´ exonuclease activity can be used to generate blunt ends from a 3´-overhang. DNA Polymerase I Large (Klenow) Fragment is purified from a recombinant E.coli strain.

Concentration & Storage Condition
- 5 unit/㎕. Store at -20℃.

Storage Buffer
- 50 mM Tris-HCl, pH 7.5, 1 mM DTT, 0.1 mM EDTA, 50% (v/v) glycerol.

10x Reaction Buffer
- 500 mM Tris-HCl, pH 7.2, 100 mM MgSO₄, 1 mM DTT.

Unit Definition
- One unit is defined as the amount of enzyme that incorporates 10 nmole of total dNTP into TCA-insoluble material in 30 min at 37°C.

The reaction conditions are: 67 mM potassium phosphate, pH 7.5, 6.7 mM MgCl₂, 1 mM DTT, 50 ㎍/ml activated calf thymus DNA and 33 μM each of dATP, dCTP, dGTP and dTTP (a mix of unlabeled and [³H]dTTP).

QC Tests
- Activity, exo and endonuclease activity test, SDS-PAGE purity, performance tests.
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