DNA Polymerase I Large (Klenow) Fragment
Fornecedor: Promega
DNA Polymerase I Large (Klenow) Fragment is a 68kDa C-terminal fragment of E. coli DNA Polymerase I that lacks the 5´→3´ exonuclease activity of intact DNA polymerase I but retains its 5´→3´ polymerase, 3´→5´ exonuclease and strand displacement activities.
- Use in a variety of molecular applications
- Heat-inactivate by heating at 75 °C for 10 minutes
- Active in many Promega 1X restriction enzyme buffers
- Supplied at a concentration of 5 U/μl
The 5´→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´→5´ exonuclease activity can be used to generate blunt ends from a 3´-overhang.
Unit definition: One unit is defined as the amount of enzyme required to catalyse the incorporation of 10 nmol of deoxynucleotide into acid-precipitable material in 30 minutes at 37 °C.
Fornecimento: Kit includes DNA Polymerase, Klenow fragment (5 U/µl) supplied in Storage Buffer: 50 mM Tris-HCl (pH 7,5), 1 mM DTT, 0,1 mM EDTA and 50% (v/v) glycerol, and with 10X Reaction Buffer: 500 mM Tris-HCl (pH 7,2), 100 mM MgSO₄ and 1 mM DTT.
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