DNA Polymerase I Large (Klenow) Fragment, Exonuclease Minus

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DNA Polymerase I Large (Klenow) Fragment, Exonuclease Minus

  • Recombinant DNA Polymerase I Large (Klenow) Fragment, Exonuclease Minus
  • Multiple applications including DNA labeling and sequencing
DNA Polymerase I Large (Klenow) Fragment, Exonuclease Minus (Klenow Fragment, Exo-) is a DNA-dependent DNA polymerase which catalyzes the 5¢¥¡æ3¢¥synthesis of DNA and lacks both the 5¢¥¡æ3¢¥ and the 3¢¥¡æ5¢¥ exonuclease activities.
 
Source:Purified from a strain of E. coli that carries a Klenow Fragment, Exo- expression plasmid.
Applications:
  • Random primed labeling (1)
  • DNA sequencing by the Sanger dideoxy method (2)
Heat inactivation: 70¡ÆC for 15 minutes
 
Storage: Enzyme and buffer should be stored at -20¡ÆC. Enzyme is stable for 12 months if handled properly.
 
Additional Information:
  • Klenow Fragment, Exonuclease Minus, will leave a single-base 3¢¥ overhang on many DNA fragments during 5¢¥-overhangs fill-in reactions. This enzyme is not recommended for preparation of blunt-ended fragments.
References:
1.  Tabor, S. and Struhl, K. (1989) In DNA-Dependent DNA Polymerases. F. M. Ausebel, R. Brent, R. E. Kingston, D. D. Moore, J. G. Seidman, J. A. Smith and K. Struhl (Eds.), Current Protocols in Molecular Biology, pp. 3.5.7-3.5.10.
2.  Sanger, F. et al. (1977) Proc. Natl. Acad. Sci. USA, 74, 5463-5467.
 
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