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T4 DNA Ligase (1 Weiss Unit/ul) Quantity in Basket: None
Code: BN19100
Price: $47.00
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/µl
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 3.3µM [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 20µl reaction mixture and a 5' DNA termini concentration of 0.12µM (300µg/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
BN19100
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