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You are here : PCR / DNA Amplification » Enzymes and chemicals for PCR » T 4 DNA Ligase

T 4 DNA Ligase

T 4 DNA Ligase high concentrated catalyzes the formation of a phosphodiester bond between juxtaposed 5´ phosphate and 3´ hydroxyl termini in duplex DNA or RNA.

Cat.-no Description Amount Price € Shop
 402-002  T 4 DNA Ligase (weiss units)      2000 units  10.00  add 
 402-010  T 4 DNA Ligase (weiss units)   10.000 units  29.00  add

T4 DNA Ligase:  Request/order by e-mail          
T 4 DNA Ligase: Datasheet
T 4 DNA Ligase: Deutsche Beschreibung

Applications:
- Cloning of restriction fragments
- joining linkers and adapters to blunt-ended DNA
- gene (gene fragments) synthesis.

Description:
T4 DNA Ligase catalyzes the formation of a phosphodiester bonds between 5' phosphate and 3' hydroxyl termini in duplex DNA/RNA. This enzyme can join blunt end and cohesive end termini, repair single stranded nicks in duplex DNA, RNA, or DNA/RNA hybrids.

Concentration: 100-200 u/µl

Source:
Isolated from E.coli strain that carries the cloned DNA ligase gene from bacteriophage T4

Usage:
For most cohesive end ligations, a 30 minute incubation at 20°C is sufficient. Incubations at 16°C for 4-16 hours are routinely used for the majority of applications.
Ligation of blunt ends and single-base pair overhang fragments requires more enzyme to achieve the same extent of ligation as cohesive end DNA fragments. Ligation may be enhanced by addition of PEG, or by reducing the rATP concentration. ATP is an essential cofactor for the reaction.

Storage buffer:
50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM DTT, 50% glycerol.

Unit definition:
One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of lambda DNA in 30 minutes at 16°C at 5' termini concentration of 0.12 µM (300 µg/ml). One Cohesive End Ligation Unit equals  0.015 Weiss units. One Weiss unit equals 67 Cohesive End Ligation Units.

Reaction buffer (10X):
500 mM Tris HCL (pH 7,8), 100 mM MgCl2, 100 mM DTT, 10 mM ATP.

Quality Assurance:
Free of contaminating exonuclease and endonuclease

Storage: shipped on blue ice, 24 months at -20°C

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Overview

Overview enzymes
and chemicals:
Uracil-DNA Glycosylase
Uracil-DNA Glycosylase (UDG) prevents carry over of DNA in PCR reactions
T 4 DNA Ligase
T 4 DNA Ligase high concentrated catalyzes the formation of a phosphodiester bond between juxtaposed 5´ phosphate and 3´ hydroxyl termini in duplex DNA or RNA.
Agarose 50bp-50kbp
Universal Agarose from GeneON is ideal for use as standard agarose for analytical as well as preparative nucleic acid electrophoresis of fragments from 50 bp to 50 kbp. Even at low concentrations the gel produced is very firm
Proteinase K
Proteinase K isolated from Tritirachium album is used for protease digestion during DNA and RNA preparation. Proteinase K from GeneON is very active and stable
IPTG
IPTG (isopropyl-beta-D-thiogalactopyranoside) is a highly stable synthetic analog of lactose. It is used in conjunction with X-Gal to determine the lac phenotype in blue/white colony screening
X-Gal
X-Gal: On combination with suitable bacterial cloning vectors, host strains,


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