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T4 DNA recombinant protein :: T4 DNA Ligase Recombinant Protein

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Catalog # MBS142427
Unit / Price
  20000 IU  /  $160 +1 FREE 8GB USB
  100000 IU  /  $345 +1 FREE 8GB USB
  500000 IU  /  $1,165 +1 FREE 8GB USB
T4 DNA recombinant protein
Product Name

T4 DNA Ligase, Recombinant Protein

Popular Item
Full Product Name

Recombinant T4 DNA Ligase

Product Synonym Names
T4 DNA; T4 DNA Ligase Recombinant; DNA ligase 4; EC 6.5.1.1; DNA ligase IV; Polydeoxyribonucleotide synthase [ATP] 4
Research Use Only
For Research Use Only. Not for use in diagnostic procedures.
Host
Escherichia Colilambda lysogen NM 989
Form/Format
(167,000 U/ml) 50mM Tris-HCl (pH 7.8 at 25°C), 10mM MgCl2, 10mM DTT, 1mM ATP, 25 ug/ml BSA and DNA (0.1 to 1 um in 5´ termini).

Optimal ligation occurs at 16°C.
Physical Appearance
Sterile filtered liquid formulation
Storage Buffer
50mM KCI, 10mM Tris-HCI (pH 7.4),0.1 mM EDTA, 1mM DTT, 200 ug/ml BSA and 50% glycerol. Store at -20°C.
Unit Definition
1. One unit is defined as the amount of enzyme required to give 50% ligation of Hind III fragments of DNA (5´ DNA termini concentration of 0.12 uM, 300- µg/ml) in a total reaction volume of 20 ul in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.

2.One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37°C.
Biological Activity
One Weiss unit is equivalent to circa 67 cohesive-end ligation units.

• T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concentration is > 200mM.
• Ligation of blunt-ended and single-base pair overhang fragments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt-end ligation may be enhanced by addition of PEG 4000 (10% w/v final concentration) or hexamine chloride, or by reducing the ATP concentration to 50uM.
• To dilute T4 DNA Ligase that will subsequently be stored at –20°C, 50% glycerol storage buffer should be used; to dilute for immediate use, 1x T4 DNA Ligase reaction buffer can be used.
Heat Inactivation
T4 DNA Ligase can be inactivated by incubation at 65°C for 10 minutes.
Quality Control
Purified free of contaminating endonucleases and exonucleases. Each lot of T4 DNA ligase is also tested in a mock cloning assay, which reveals any damage to the ligated DNA termini. Greater than 99.9% of the termini remain undamaged in this assay.


Exonuclease Activity:

Incubation of a 50µl reaction containing 13,000 units of T4 DNA Ligase with 1µg of a mixture of single and double-stranded [3H] E. coli DNA (200,000 cpm/ug) for 4 hours at 37°C released < 0.3% of the total radioactivity.


Endonuclease Activity:

Incubation of a 50µl reaction containing 13,000 units of T4 DNA Ligase with 1µg of X174 RF I DNA for 4 hours at 37°C resulted in < 5% conversion to RFII as determined by agarose gel electrophoresis.


Nuclease Activity :

Incubation of 13,000 units for 18 hours in assay buffer (without ATP) with Hind III fragments of gamma DNA yielded a clear and sharp banding pattern on agarose gels.
Preparation and Storage
Two years when stored at -20°C, 2 weeks at 4°C.
Other Notes
Small volumes of T4 DNA recombinant protein vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.
Related Product Information for
T4 DNA recombinant protein
Description: T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5' -phosphate and 3' -hydroxyl termini in duplex DNA or RNA. This enzyme will join blunt end and cohesive end termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.
Application Notes for T4 DNA recombinant protein
Cloning of restriction fragments.

Joining linkers and adapters to blunt-ended DNA.
Precautions
All of MyBioSource's Products are for scientific laboratory research purposes and are not for diagnostic, therapeutics, prophylactic or in vivo use. Through your purchase, you expressly represent and warrant to MyBioSource that you will properly test and use any Products purchased from MyBioSource in accordance with industry standards. MyBioSource and its authorized distributors reserve the right to refuse to process any order where we reasonably believe that the intended use will fall outside of our acceptable guidelines.
Disclaimer
While every efforts were made to ensure the accuracy of the information provided in this datasheet, MyBioSource will not be liable for any omissions or errors contained herein. MyBioSource reserves the right to make changes to this datasheet at any time without prior notice.

It is the responsibility of the customer to report product performance issues to MyBioSource within 30 days of receipt of the product. Please visit our Terms & Conditions page for more information.
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