Thermo Scientific™ T4 RNA Ligase (10 U/μL)

Catalog No.	Quantity
FEREL0021	1,000 units

Catalog No. FEREL0021 Supplier Thermo Scientific™ Supplier No. EL0021

Description

Catalyze the ATP-dependent formation of phosphodiester bonds between 5'-P and 3'-OH termini of oligonucleotides, single-stranded RNA and DNA.

T4 RNA Ligase catalyzes the ATP-dependent intra- and intermolecular formation of phosphodiester bonds between 5'-phosphate and 3'-hydroxyl termini of oligonucleotides, single-stranded RNA and DNA.

The minimal substrate is a nucleoside 3',5'-biphosphate in intermolecular reaction and oligonucleotide of 8bases in intramolecular reaction.

• Source: E. coli cells with a cloned gene 63 of bacteriophage T4
• Molecular Weight: 43.6 kDa monomer

Quality Control:

• The absence of ribonucleases, exodeoxyribonucleases, endodeoxyribonucleases and phosphatases confirmed by appropriate quality tests.

Source:

• E.coli cells with a cloned gene 63 of bacteriophage T4

Molecular Weight:

• 43.6kDa monomer

Definition of Activity Unit:

• One unit of the enzyme catalyzes the conversion of 1nmol of 5ft.-[³²P]-(A)_12-18 to a phosphatase-resistant form in 30 min. at 37°C
• Enzyme activity is assayed in the following mixture: 50mM Tris-HCl (pH 7.5), 10mM MgCl₂, 10mM DTT, 1mM ATP, 10μM 5'-[³²P]-(A)_12-18 (10μM in 5ft.-termini)

Storage Buffer:

• The enzyme is supplied in:20mM Tris-HCl (pH 7.5), 1mM DTT, 50mM KCl, 0.1mM EDTA, 0.03% (v/v) ELUGENT Detergent and 50% (v/v) glycerol

10X Reaction Buffer:

• 500mM Tris-HCl (pH 7.5 at 25°C), 100mM MgCl₂, 100mM DTT, 10 mM ATP

Inhibition and Inactivation:

• Inhibitors: metal chelators, SH group-modifying reagents (8).
• Inactivated by heating at 70°C for 10min.

Recommended for:

RNA 3'-end labeling with cytidine 3',5'-bis [alpha-³²P] phosphate (1); Joining RNA to RNA (2); Synthesis of oligoribonucleotides and oligodeoxyribonucleotides (3, 4); Specific modifications of tRNAs (5); Oligodeoxyribonucleotide ligation to single-stranded cDNAs for 5ft. RACE (Rapid Amplification of cDNA Ends) (6); Site-specific generation of composite primers for PCR (7)

Note:

The recommended BSA concentration in the reaction mixture is 0.1mg/ml.

This product(s) resides on a Fisher Scientific GSA or VA contract. If you are viewing this page as a nonregistered user, the price(s) displayed is List Price. To view your GSA or VA contract pricing, log in using your account number, or become a registered user by contacting one of our Customer Service teams. You can also view your contract price by searching for this item(s) on GSA Advantage. To place an order, contact Fisher Scientific Customer Service.

Specifications

• Concentration: 10 U/μL
• Enzyme: T4 RNA Ligase
• Compatible Buffer: 10X Reaction Buffer
• Quantity: 1,000 units
• Product Type: T4 RNA Ligase

Frequently Asked Questions (FAQs)

What are some of the problems associated with sticky-end cloning?

The amplified DNA needs to be purified from the PCR mixture components prior to cloning. The dNTPs carried over from the PCR are competitive inhibitors for ATP in the ligation reaction.

If during synthesis of the PCR primers their chemical integrity has been compromised by either a base substitution or modification, the enzyme recognition site may in actuality not exist. If this is the case, PCR products will be resistant to digestion with restriction enzymes. It may be necessary to use a higher concentration of the restriction enzyme and to incubate at the appropriate temperature overnight to ensure cutting.

For Research Use Only. Not for use in diagnostic procedures.