Pages
Products

T4 DNA Ligase

For research use only. Not intended for any clinical use.
Cat.No.
EMQZ1404
Description
T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5' phosphate and 3' hydroxyl termini in duplex DNA or RNA. 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.
Applications
• Cloning of restriction fragments• Joining linkers and adapters to blunt-ended DNA
Concentration
400,000 U/ml
Reaction Conditions
1×T4 DNA Ligase Reaction Buffer [50 mM Tris-HCl (pH 7.5 at 25°C), 10 mM MgCl2, 10 mM DTT, 1 mM ATP; incubate at 16°C.
Size/Form
20,000 U; 100,000 U
Source
Purified from a recombinant E. coli strain.
Storage
Store at -20°C
Unit Definition
One unit is the amount of enzyme required to yield 50% ligation of HindIII fragments of lambda DNA in 30 min at 16 °C in a 20 μl assay mixture containing a DNA terminus concentration of 0.12 μM.

Publications

Q & A

Customer Reviews

Customer Q&As
Can sticky ends of DNA be ligated?

A: The enzyme can ligate both blunt and sticky ends of DNA.

Can this enzyme only ligate DNA ends?

A: The T4 DNA ligase enzyme catalyzes the formation of phosphodiester bonds between adjacent 5' phosphate and 3' hydroxyl ends in double-stranded DNA or RNA, and repairs single-strand breaks in double-stranded DNA, RNA, or DNA/RNA hybrids.

What are the applications of this enzyme?

A: 1. cloning of DNA fragments generated by restriction endonucleases. 2. cloning of PCR products. 3. linking double-stranded oligonucleotide linkers or connectors to DNA. 4. targeted mutagenesis. 5. amplified fragment length polymorphism (AFLP). 6. ligase-mediated RNA detection. 7. nick repair in double-helical DNA, RNA, or DNA/RNA hybrids. 8. autocycling of linear DNA.

What should be noted when using this enzyme?

A: ?The binding of T4 DNA ligase to DNA may result in band displacement in the agarose gel. To avoid this, incubate samples at 70°C for 5 minutes or at 65°C for 10 minutes with 6X DNA loading dye & SDS solution and cool on ice before electrophoresis.

Does the ligand product using this enzyme allow for direct electrotransfer?

A: It is not recommended that the T4 DNA ligase be removed from the ligation mixture prior to electrotransformation, preferably using a centrifugation column or chloroform extraction. The extracted DNA can be further precipitated using ethanol and then electrotransformed to improve transformation efficiency.

Ask a Question

If your question is not addressed through these resources, you can fill out the online form below and we will answer your question as soon as possible.

Customer Reviews
Can act on both DNA and RNA

This enzyme catalyzes the formation of phosphodiester bonds in double-stranded DNA or RNA and repairs single-stranded cuts in double-stranded DNA, RNA or DNA/RNA hybrids.

United States

12/19/2022

Faster reaction

The enzyme completes the ligation of sticky ends within 10 minutes at room temperature and is supplied in PEG solution for efficient flat end ligation.

United States

12/20/2022

Active in a variety of reaction systems

Active in restriction endonuclease, PCR and RT buffer (when supplemented with ATP).

United States

01/27/2023

Write a Review

Write a review of your use of Biogene products and services in your research. Your review can help your fellow researchers make informed purchasing decisions.

Needs improvement

Satisfaction

General satisfaction

Very satisfaction

CBpromise

Our promise to you:
Guaranteed product quality, expert customer support.

24x7 CUSTOMER SERVICE
CONTACT US TO ORDER
Quick Inquiry

Inquiry