T4 DNA Ligase with reaction buffer and ATP

Product#: 02-050
$35.00

Size:

  • 20000 Units
  • 5x 20000 Units
Availability:
Ships in 1-2 weeks

T4 DNA Ligase with reaction buffer and ATP

Cat. No. 02-050
Store at -20°C
Size 20000 Units/ 20000 Units x5

Description

Bacteriophage T4 derived DNA ligase catalyzes the formation of phosphodiester bonds between 3’-OH termini and 5’-P termini in duplex DNA or RNA (1). This enzyme will join blunt end and cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids。 T4 DNA ligase was expressed in E.coli in large quantities and highly purified. MW is 55.3 kDa.

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Fig.1 SDS-PAGE of T4 DNA ligase protein


 

Fig.2 DNA ligation activity. Ligation of Hind III fragments of λ DNA using 1 unit of T4 DNA ligase Incubation at 16℃ for 0, 10, 20, 30, and 60 min. 

Applications

  1. Insertion of DNA fragment into a vector
  2. Linker (or Adaptor) ligation with DNA fragment

Storage conditions

10mM Tris-HCl (pH 7.6), 50mM KCl, 0.1mM EDTA, 1mM dithiothreitol, 50% glycerol Store at -20℃

Concentration

400 U/ul, where one unit is the amount of enzyme that ligates more than 90% of 6 ug of λDNA-HindIII fragments in a 20μl mixture in 30 minutes at 16℃.

Quality Assurance

Greater than 95% protein determined by SDS-PAGE (CBB staining) The absence of endonucleases and exonucleases was confirmed.

Reagents Supplied with Enzyme

10 x T4 Ligase Reaction Buffer (T4-Lig): 500mM Tris-HCl (pH 7.6), 100mM MgCl2, 10 mM ATP, 100mM dithiothreitol 

Data Link

UniProtKB/Swiss-Prot P00970 (DNLI_BPT4) 

References

  1. Weiss B et al (1968) “Enzymatic breakage and joining of deoxyribonucleic acid.” J. Biol. Chem. 243: 4543-4555 PMID: 4879167
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