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T4 DNA Ligase

Item #: ASCDNART8 

T4 DNA Ligase

Intact Genomics T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA.  This enzyme joins DNA fragments with either cohesive or blunt termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.
 

Quality Control 
Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

Intact Genomics T4 DNA Ligase displays up to 3-5X higher ligation efficiency than the nearest competitor.

Product Source
E. coli strain expressing a recombinant clone

Quality Control 
Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

Contents & Storage

  1. T4 DNA Ligase
  2. 10x T4 DNA Ligase Reaction Buffer (w/o ATP)
  3. 10 mM ATP

Store all contents at -20 °C.

Storage Buffer 
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25 °C

10x T4 DNA Ligase Reaction Buffer (w/o ATP) 
500 mM Tris-HCl, 100 mM MgCl, 100  mM DTT, pH 7.5 @ 25 °C

Note
10x T4 DNA ligase buffer does not contain ATP. You need to add ATP separately.

Unit Definition 
One Weiss unit is defined as the amount of enzyme required to convert 1 nmol of 32P from pyrophosphate into Norit-absorbance material in 20 minutes under standard assay conditions.

Protocol

  1. Set up reaction buffer in a microcentrifuge tube on ice. Use a molar ratio of 1:3 vector to insert DNA.

  


Component

10 µl Reaction

Vector DNA

x µl

Insert DNA

x µl

10 mM ATP

1.0µl

10x T4 Ligase Buffer

1.0µl

T4 DNA Ligase

1.0µl

Add H2O up to

10.0µl

  

  1. Gently mix the reaction and centrifuge briefly.
  2. For cohesive ends, incubate 16 °C for overnight or at room temperature for 30 min.
  3. For blunt ends, incubate 16 °C for overnight or at room temperature for 2 hrs.
  4. Heat inactivate at 70 °C for 15 min.
  5. Cool on ice and transform 2 µl of the reaction into 50 µl competent cells.

PNCDNART

Pkg
No dye
 
SKU Number Pkg Vol Price Quantity Add to Cart
3212 No dye 500 Weiss units, 250 uL
428.57
Each
3216 No dye 500 Weiss units, 50 uL
428.57
Each
3217 No dye 2,000 Weiss units, 200 uL
714.28
Each
T4 DNA Ligase
T4 DNA Ligase
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