T4 DNA Ligase
recombinant form of the enzyme from T4 phage, solution
For further processing only.
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T4 DNA Ligase material number and pack size: Material Number Pack Size 10909246103 custom fill
The enzyme is supplied without reaction buffer.
- Use T4 DNA Ligase for ligation of DNA fragments. (5 x 103 U/ml)
- Use T4 DNA Ligase to ligate DNA fragments with blunt or overlapping ends.
Enzyme activities: T4 DNA Ligase catalyzes the formation of phosphodiester bonds between neighbouring 3'-hydroxyl and 5'-phosphate ends in double-stranded DNA. Sticky- and blunt-ended DNA fragments are ligated. Single-stranded nicks in double-stranded DNA are also closed.
Appropriate ligation buffer, 10x concentrated: Tris/HCl, 660 mmol/L; MgCl2, 50 mmol/L; DTT, 50 mmol/L; ATP, 10 mmol/L, pH 7.5 at +20°C (Note: ATP is not stable).
pH optimum: 7.2-7.8
Divalent ion requirement: Mg2+
Inactivation: After 10 minutes heat denaturation at +65°C ligase activity is stopped. Appearance: Clear, colorless solution
Storage buffer: Tris/HCl, 20 mmol/L; KCl, 60 mmol/L; DTE, 5 mmol/L; EDTA, 1 mmol/L; glycerol, 50% (v/v); pH approximately 7.5 at +4°C
Volume activity: ≥5 U/μL
Unit definition: One unit T4 DNA Ligase is defined as the amount of enzyme which converts 1 nmol of [32P] from pyrophosphate into Norit-absorbable material in 20 minutes at +37°C.
Glycosylases (M13mp11(U) ssDNA): Not detectable in up to 10 U after 16 hours incubation at +37°C.
Nicking activity (pBR322 DNA): Not detectable in up to 10 U after 16 hours incubation at +37°C.
Animal-derived additives: None
Stability: At -15 to -25°C within specification range for 18 months.