E. coli DNA Ligase
Lig
10 U/μL
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- Application
- Ligation (Lig)
- Characteristics
- E.coli DNA Ligase is an NAD+-dependent enzyme that catalyzes the formation of a phosphodiester bond between cohesive 3'-hydroxyl and 5'-phosphoryl termini of double-stranded DNA (dsDNA). This enzyme is also active on nicked DNA but is not effective for the formation of DNA-RNA or RNA-RNA hybrids.
- Components
- Enzyme supplied with 10X Reaction Buffer
- Unit Definition
- One unit is defined as the amount of E. coli DNA Ligase that is required to give 50% ligation of HindIII-digested λ DNA in a total reaction volume of 20 μl in 30 minutes at 16°C in 1X E.coli DNA Ligase Reaction Buffer (concentration of 5' DNA termini is 0.12 μM [300 μg/ml]).
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- Comment
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- Ligation of dsDNA with cohesive termini
- cDNA cloning of products from second strand cDNA synthesis experiments
- Alternative to T4 DNA Ligase when bluntend ligation is not required
- For ligation of bluntend fragments use T4 DNA Ligase (abm Cat. No. G467)
- Restrictions
- For Research Use only
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- Concentration
- 10 U/μL
- Buffer
- 50 mM Tris-HCl ( pH 7.5), 1 mM EDTA, 1 mM DTT, 200 mM NaCl, and 50 % (v/v) Glycerol.
- Storage
- -20 °C
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