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T4 DNA Ligase(Fast) is produced by E.coli carrying T4 bacteriophage gene 30. The enzyme catalyzes the formation of phosphodiester bonds between 5′-phosphate groups and 3′-hydroxyl groups of double-stranded DNA or RNA. It can repair single-strand nicks in double-stranded DNA, RNA, or DNA/RNA hybrids and can ligate DNA fragments with sticky or blunt ends. It is not active on single-stranded nucleic acids. The enzyme is primarily used for cloning of restriction endonuclease-digested DNA fragments, site-directed mutagenesis, cloning of PCR products, linear DNA circularization, and repair of double-stranded DNA nicks. T4 DNA Ligase (Fast) requires ATP as a cofactor and completes sticky end ligation reactions at room temperature in just 10 minutes.
Components
| Component | Amount |
|---|---|
| T4 DNA Ligase(Fast) (5 U/μl) | 200 μl |
| 10×T4 DNA Ligase Buffer | 2×1 ml |
| 50% PEG | 1 ml |
Note: 1 U=1 Weiss unit.
Storage Condition
Store at -20℃ for 2 years.
