T4 DNA Ligase

Downloads

Certificate of Analysis PDF -Current Lot
MSDS PDF -Current Lot    

Reagents Supplied

5X T4 DNA Ligase Buffer

Source

T4 bacteriophage of Escherichia coli

Applications

• Catalyzes the joining of duplex DNA molecules at blunt ends (1)
• Covalently joins DNA fragments with complementary cohesive ends
• Seals single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids
• Suitable for cloning of restriction fragments and joining linkers or adapters to blunt-ended DNA (2)
• Ultrapure recombinant enzyme

Unit Definition

One unit is the amount of enzyme required to convert 1 nmol of 32P from pyrophosphate into Norit-adsorbable form in 20 minutes at 37°C
Note: 67 cohesive end ligation units are the equivalent of one Weiss unit

Heat Inactivation

65ºC for 15 minutes

Storage Buffer

10 mM Tris-HCl (pH 7.5 at 22°C)
50 mM KCl
1 mM dithiothreitol
50% (v/v) glycerol

Assay Conditions

66 mM Tris-HCl (pH 7.6 at 22°C)
6.7 mM MgCl₂
10 mM dithiothreitol
67 µM ATP
3.3 µM [α-³²P]Na₄P₂O₇ (1X108 CPM/µmol)
Reaction volume of 30 µl

Storage Conditions

Store at -20°C
Product shipped on dry ice

References

(1) Weiss, B., Jacquemin-Sablan, A., Live, T.R., Fareed, G.C. and Richardson, C.C. (1968) J. Biol. Chem. 243, 4543-4555
(2) Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, 2nd Edition, 1.53-1.73