Thermo Fisher EL0011 1,000 units T4 DNALigase (5 U/μL)
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EL0011 1,000 unitsT4 DNA Ligase (5 U/μL)

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Thermo Scientific T4 DNA Ligase catalyzes the connection between the side-by-side 5-phosphate and 3-hydroxyl ends of double helix DNA or RNA Formation of phosphodiester bonds. The enzyme can repair single-stranded gaps in double-helix DNA, RNA, or DNA/RNA hybrids. It can also bind DNA fragments with sticky or blunt ends, but is inactive against single-stranded nucleic acids.

T4 DNA ligase requires ATP as a cofactor.

Product Advantages

• Active in Themo Scientific restriction enzymes, PCR and RT buffers (when supplemented with ATP)
• Fast —Cohesive end ligation in 10 minutes at room temperature
• Supplied with PEG solution for efficient blunt-end ligation

Applications

• Cloning DNA fragments generated by restriction enzymes
•Clone PCR products
• Attach double-stranded oligonucleotide linkers or adapters to DNA
• Site-directed mutagenesis
• Amplify fragment length polymorphisms (AFLP)
• Ligase-mediated RNA detection (seeReference 3)
• Gap repair in duplex DNA, RNA, or DNA/RNA hybrids
• Self-circularization of linear DNA.

Includes

• T4 DNA Ligase
• 10X T4 DNA Ligase Buffer
• 50% PEG Solution

Notes

• Binding of T4 DNA ligase to DNA may cause band shifts in agarose gels. To avoid this, use 6X DNA Loading Dye & SDS Solution at 70°C Incubate samples for 5 minutes at 65°C or 10 minutes at 65°C, cool on ice before electrophoresis.
•During the transformation process, the volume of the ligation reaction mixture should not exceed 10% of the volume of the competent cells.
•Before electroporation, remove T4 DNA ligase from the ligation mixture using spin columns or chloroform extraction. The extracted DNA can be further ethanol precipitated.
For Research Use Only. Not for use in diagnostic procedures.