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

Name: T4 DNA Ligase (5 U/μL), 1,000 units
Brand: Thermo Fisher
SKU: EL0011

Quantity: 1, 000 units, Unit Size: Each, Catalyze the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA with the Thermo Scientific™ T4 DNA Ligase. The enzyme repairs single-strand nicks in duplex DNA, RNA, or DNA/RNA hybrids. It also joins DNA fragments with either cohesive or blunt termini, but has no activity on single-stranded nucleic acids.The T4 DNA Ligase requires ATP as a cofactor.

Catalog Number: EL0011

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Category

DNA Ligases

Pack Size000 units / pack

TagsModifying Enzymes Enzymes and Inhibitors

CollectionsThermo Fisher Enzymes & Inhibitors for PCR, Cloning & Protein Protection Thermo Fisher Clinical Analyzers & Instruments for Diagnostic Labs Thermo Fisher General Lab Supplies for Daily Workflows — Nalgene, Nunc, Samco & Matrix

Description

Quantity: 1,000 units
Unit Size: Each
Highlights
• Active in Themo Scientific restriction enzyme, PCR, and RT buffers (when supplemented with ATP)
• Fast – sticky-end ligation is completed in 10 minutes at room temperature
• Supplied with PEG solution for efficient blunt-end ligationApplications
• Cloning of restriction enzyme generated DNA fragments
• Cloning of PCR products
• Joining of double-stranded oligonucleotide linkers or adaptors to DNA
• Site-directed mutagenesis
• Amplified fragment length polymorphism (AFLP)
• Ligase-mediated RNA detection (seeReference 3)
• Nick repair in duplex DNA, RNA or DNA/RNA hybrids
• Self-circularization of linear DNA.Notes
• Binding of T4 DNA Ligase to DNA may result in a band shift in agarose gels. To avoid this, incubate samples with 6X DNA Loading Dye & SDS Solution at 70°C for 5 min or 65°C for 10 minutes and chill on ice prior to electrophoresis.
• The volume of the ligation reaction mixture should not exceed 10% of the competent cell volume in the transformation process.
• Prior to electro-transformation, remove T4 DNA Ligase from the ligation mixture by spin column or chloroform extraction. The extracted DNA can be further precipitated with ethanol.

Specifications

General

Compatible Buffer	10X T4 DNA Ligase Buffer
Product Type	T4 DNA Ligase
Quantity	1,000 units
Concentration	5 U/μL
Enzyme	DNA Ligase
Unit Size	Each

FAQs

Question: What enzyme concentration and total activity are provided?

Answer:The product supplies 1,000 units of T4 DNA ligase at 5 U/μL.

Question: What reaction components are included?

Answer:It includes the enzyme, 10X ligase buffer and 50% PEG solution.

Question: What cofactor is required for ligation?

Answer:ATP is required.

Application Note

T4 DNA Ligase (5 U/μL), 1,000 units is designed for ligation of duplex DNA fragments during cloning and related nucleic acid manipulation. The enzyme catalyzes formation of phosphodiester bonds between adjacent 5′-phosphate and 3′-hydroxyl groups, joining compatible DNA ends and repairing single-strand nicks in duplex substrates. It can ligate cohesive-ended or blunt-ended DNA and also supports nick repair in duplex DNA, RNA and DNA/RNA hybrids; it does not ligate isolated single-stranded nucleic acids. ATP is required as a cofactor. The package provides 1,000 enzyme units at a concentration of 5 U/μL and includes 10X T4 DNA Ligase Buffer and 50% PEG solution. Cohesive-end ligation can be completed in approximately ten minutes at room temperature under suitable conditions, while PEG can support efficient blunt-end ligation. Common applications include cloning restriction fragments or PCR products, adding double-stranded linkers or adapters, site-directed mutagenesis and recircularizing linear DNA molecules. The inclusion of buffer and PEG allows the enzyme to be used for both compatible-end and blunt-end ligation without separately sourcing these supporting reagents.

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