Navigation Links
Ligase, T4 DNA from USB Corp.

ProductsLigase, T4 DNA from USB Corp.
Company USB Corp.
Item Ligase, T4 DNA
Price 
Description Source: E. coli strain containing an overproducing clone of T4 DNA Ligase.
Description:
T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed
5' phosphoryl and 3'-hydroxyl termini in duplex DNA or RNA. It repairs single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids and will join both blunt-end and cohesive-end fragments of duplex DNA or RNA(1).
Tested User Friendly
Properties:
Molecular Weight: 55,000 Da
Optimum pH: 7.5 - 7.8
Optimum Temperature: 16C for exchange reaction
Requirements for Divalent Cation: Mg2+, Mn2+
Optimum Mg2+ Concentration: 10mM
Sulfhydryl Requirement: 2-mercaptoethanol, dithiothreitol
Stimulators: Spermine (0.5 to 1mM), Spermidine (0.5 to 1mM)
Inhibitors: Na (>0.2M), K (>0.2M)
Inactivation: Heat at 65C for 10 minutes

Purity:
Greater than 95% pure as determined by SDS-PAGE. Tested for contaminating endonucleases, exonucleases and ribonucleases.
Storage Buffer:
25mM Tris-HCl (pH 7.6), 100mM NaCl, 1mM DTT, 0.5mM EDTA, 50% glycerol.
Shipping and Storage:
Shipped on dry ice. Store at -20C.
Assay Conditions:
T4 DNA Ligase is assayed according to the pyrophosphate exchange method of Weiss
et al. The reaction mixture (300 l) contains 67mM Tris-HCl, (pH 7.8), 6.7mM MgCl2, 10mM DTT, 66M ATP and 3.3M radiolabeled pyrophosphate. Incubation is at 37C for
20 min.

Note: ATP is an essential cofactor for the reaction. This contrasts with E. coli DNA Ligase which requires NAD.
Unit Definition:
One Weiss unit is the amount of enzyme required to convert 1 nmol of radiolabeled phosphate from pyrophosphate into absorbable material in 20 min at 37C under standard assay conditions. One Weiss unit equals about 67 cohesive end ligation units.
Concentration:
PN 70005: 1 unit/l; PN 70042: greater than or equal to 5 units/l
Tested User FriendlyTM Functional Test:
Re-ligation of linearized plasmid DNA >75% of control, as determined by counting transformed bacterial colonies.

Applications:

*Ligation of cohesive-end or blunt-end termini.
*Ligation of synthetic linkers or adaptors.

PROTOCOL FOR DNA LIGATION
For each 10 l of ligation reaction, a combination of 2040 ng of vector DNA and a 1:3-10 fold molar excess of foreign DNA will produce an adequate yield of recombinant genomes for
most cloning purposes.
1. Combine the following:
Vector DNA (2040 ng) __ l
Insert DNA (310-fold molar excess) __ l
Ligation Buffer, 10X 5 l
Water __ l
T4 DNA Ligase __ l (1 unit)

Total 50 l
2. Mix and incubate at 16C for 16 hrs.

Note: Km for the activity of T4 DNA Ligase on blunt-ended DNA is nearly 100 times higher than that on DNA with cohesive ends. Therefore, ligation of blunt-ended DNA should be carried out at a higher concentration of both vector and insert DNA.

To stimulate the blunt-end ligation reaction, 150200mM NaCl and 5% PEG can be added.
References:

*ROSSI, R., MONTECUCCO, A., CIARROCHI, G. AND BIAMONTI, G. (1997) Nucl. Acids Res. 25 (11), 2106-2113.
*WEISS, B., JACQUEMIN-SABLON, A., LIVE, T. R., FAREED, G. C. AND RICHARDSON, C. C. (1968) J. Biol. Chem. 243, 4543-4555.

Functionally Tested 10X T4 DNA Ligase Reaction Buffer (1 ml included, PN 70087): 660mM Tris-HCl (pH 7.6), 66mM MgCl2, 100mM DTT, 660M ATP

T4 DNA Ligase Dilution Buffer (1 ml included, PN 70108): 20mM Tris-HCl (pH 7.6), 60mM KCl, 5mM DTT, 1mM EDTA, 50% glycerol
Info USB Corp.USB Corp.
26111 Miles Road
Cleveland Ohio 44128


Call USB Corp. to buy products (US and Canada only; other locations use numbers below)
USA Canada 1-216-245-1829
Customer Service: 216.765.5000
Fax Number: 216.464.5075
Web Site: http://www.usbweb.com
NOTE: Price information is approximate list price and actual prices may vary.

Related biology products :

1. Tfi DNA Ligase, 2,000U, thermostable from Bioneer, Inc.
2. S-Acetyl Coenzyme A Lithium Salt from USB Corp.
3. Mut S from USB Corp.
4. Thermo Sequenase Dye Primer Manual Cycle Sequencing Kit from USB Corp.
5. Phenol Red, Free Acid from USB Corp.
6. Dithiothreitol from USB Corp.
7. Bromophenol Blue from USB Corp.
8. Xylene Cyanol FF from USB Corp.
9. Human Topoisomerase II, Alpha from USB Corp.
10. Calf Topoisomerase I from USB Corp.
11. M-MLV Reverse Transcriptase from USB Corp.
Rabbit polyclonal to NCoR2, prediluted ( Abpromise for all tested applications). entrezGeneID: 9612 SwissProtID: Q9Y618...
Rabbit polyclonal to TRPM8 (rating: ****) ( Abpromise for all tested applications). entrezGeneID: 79054 SwissProtID: Q3YFM7...
Sheep polyclonal to Trenbolone ( Abpromise for all tested applications). Antigen: Chemical / Small Molecule: 17 alpha Trenbolone(3)-BTG....
DNA fragmentation factor & Inhibitor of CAD...
Biology Products:
(Date:3/30/2017)... 2017  On April 6-7, 2017, Sequencing.com will host ... hackathon at Microsoft,s headquarters in Redmond, ... on developing health and wellness apps that provide a ... Genome is the first hackathon for personal genomics ... companies in the genomics, tech and health industries are ...
(Date:3/27/2017)... CENTRE, N.Y. , March 27, 2017 /PRNewswire-USNewswire/ ... Healthcare Information and Management Systems Society (HIMSS) Analytics ... Outpatient EMR Adoption Model sm . In addition, ... 12% of U.S. hospitals using an electronic medical ... CHS for its high level of EMR usage ...
(Date:3/22/2017)... 2017   Neurotechnology , a provider of ... announced the release of the SentiVeillance 6.0 ... facial recognition using up to 10 surveillance, security ... The new version uses deep neural-network-based facial detection ... utilizes a Graphing Processing Unit (GPU) for enhanced ...
Breaking Biology News(10 mins):
(Date:4/25/2017)... ... , ... Leaders of Quorum Review IRB and Kinetiq , ... this week’s Association of Clinical Research Professionals (ACRP) 2017 Meeting & Expo ... "We are excited to present subject matter expertise on topics that impact the global ...
(Date:4/24/2017)...  Dante Labs announced today the offer of whole genome ... While American individuals have been able to access WGS at ... WGS below EUR 1,000. The sequencing includes bioinformatics ... to make informed decisions about disease monitoring, prevention, nutrition, exercise, ... ...
(Date:4/21/2017)... ... ... Having worked on the design of the innovative Triton™ lab bench ... design architects from around the country at the Lab Design Conference 2017 in Raleigh, ... Casey will be at the show, where they will highlight the unique elements of ...
(Date:4/21/2017)... BELLINGHAM, Washington, USA (PRWEB) , ... April 21, ... ... for sensing, imaging, and related applications were the focus of researchers, engineers, product ... Sensing 2017 in Anaheim. , Sponsored by SPIE, the international society ...
Breaking Biology Technology: