What is T4 RNA ligase?
T4 RNA Ligase catalyzes the ATP-dependent ligation of single-stranded RNA or DNA onto the 5´-phosphoryl termini of single-stranded RNA or DNA. The enzyme, purified from recombinant E. T4 RNA Ligase is provided with 10X Reaction Buffer: 500mM Tris-HCl (pH 7.8 at 25°C), 100mM MgCl2, 50mM DTT, 10mM ATP.
What does T4 DNA Ligase require?
T4 DNA Ligase requires ATP as a cofactor. Self-circularization of linear DNA. Binding of T4 DNA Ligase to DNA may result in a band shift in agarose gels.
How does T4 RNA ligase work?
T4 RNA Ligase catalyzes the formation of phosphodiester bonds between singlestranded nucleic acids with 5′-phosphate and 3′-hydroxyl termini. The enzyme requires ATP for activity. The rate of activity is highest for RNA-RNA ligation, lower for RNA-DNA ligation and very low for DNA-DNA ligation.
What is E coli DNA ligase?
E. coli DNA Ligase is a ligation enzyme that can be used to join DNA fragments by catalyzing the formation of phosphodiester bonds between double-stranded DNA fragments containing juxtaposed 5′-phosphate termini and 3′-hydroxyl termini in the presence of the NAD cofactor.
Why T4 ligase is used in Rdna?
In Vitro ligation using T4 DNA ligase: The T4 DNA ligase can ligate the sticky ends of dsDNA very efficiently, therefore, we are using it in recombinant DNA experiment or cloning methods. Our selected vectors also have the sticky ends complementary to our digested DNA. However, the plasmid ends can be self-ligated.
What does the ligase do in DNA replication?
The ligase joins the two fragments of DNA to form a longer strand of DNA by “pasting” them together.
What is the function of DNA ligase in DNA replication?
DNA ligase joins the Okazaki fragments together into a single DNA molecule. Helicase opens up the DNA at the replication fork. Single-strand binding proteins coat the DNA around the replication fork to prevent rewinding of the DNA. Topoisomerase works at the region ahead of the replication fork to prevent supercoiling.