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T4 DNA Ligase (1 Weiss Unit/ul)
 
  Quantity in Basket: None
Code: BN19100
Price: $47.00
 
 
Quantity:
 
Untitled Document Size: 200 Weiss Units


DESCRIPTION
Purified from an E. coli strain carrying a T4 DNA ligase overproducing plamid. T4 DNA Ligase catalyzes the formation of a phosphodiester linkage between 5'-phosphoryl group and adjacent 3'-hydroxyl group of duplex DNA or RNA in a blunt-ended or cohesive-ended configuration. ATP is required for the reaction.

APPLICATIONS:

  • Gene cloning : Joining double-stranded DNA molecules with cohesive or blunt ends


  • Adaptor ligation

SOURCE
T4 bacterial phage

CONCENTRATION
1 Weiss Unit / ul
20-40 units/l
10X reaction buffer
500 mM Tris-HCl (pH 7.6), 66 mM MgCl2, 100 mM DTT, 5 mM ATP

Unit (U) Definition: One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1nmole of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37?. The reaction conditions for Weiss unit definition assay are : 66mM Tris-HCl (pH 7.6), 6mM MgCl2, 1mM DTT, 0.066mM ATP and 3.3M [32PPi]. One Weiss unit is equivalent to 67 cohesive end ligation unit which is the amount of enzyme required to give 50% ligation of Lambda DNA Hind III fragments in 30 minutes at 16? in 20l reaction mixture and a 5' DNA termini concentration of 0.12M (300g/ml).


STORAGE CONDITION
-20 °C


FEATURES
Intramolecular reation works efficiently when molar ratio of plasmid and insert DNA is equimolar and total DNA concentration is lower than 10ug/ul. For blunt-ended ligation, high concentration (~3ul) of T4 DNA ligase and blunt-ended termini are required. Adding 10 % PEG6000 and reaction at 20? increase ligation efficiency.



ORDERING INFORMATION

Catalog #
Products
Size
BN19100
T4 DNA Ligase (1 Weiss Unit/ul)
200 WU


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