TurboLigase™ kit, 500 Reactions (L0110M) ACGC80002

TurboLigase™ kit, 500 Reactions (L0110M) Molecular Biology Accelagen Inc. ACGC80002
Supplier: Accelagen Inc.
Product Code: ACGC80002
Availability: In Stock
$525.00 $475.00
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TurboLigationTM Kit enables quick ligation of both blunt-end or cohesive-end DNA fragments in just 5 minutes at room temperature. The kit combines TurboLigaseTM, an ultra-pure recombinant T4 DNA Ligase, and 5X TurboLigation Buffer, an optimized quick ligation buffer. A 5 minute TurboLigation reaction offers the same efficiency as a traditional 16oC overnight ligation reaction. The reaction product is ready for bacterial transformation without any further treatment. 

Applications:

  • Adding linkers or adapters to DNA
  • Blunt or cohesive end ligation of duplex DNA
  • DNA cloning
     

Components:
TurboLigaseTM: 5 unit/µl in Storage Buffer of 10 mM Tris-HCl, pH 7.5, 50 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, and 50% glycerol
5X TurboLigation Buffer: 250 mM Tris-HCl, pH 7.5, 50 mM MgCl2, 5 mM dithiothreitol, 5 mM ATP, 25% polyethylene glycol 8000

Stability and Storage:
TurboLigase and 5X TurboLigation Buffer are stable at -20oC for 12 months from date of receipt. Store TurboLigase and 5X TurboLigation Buffer at -20oC.
Shipped in gel packs.

TurboLigaseTM Unit Definition:
0.015 Weiss unit (1 cohesive end ligation unit) is defined as the amount of TurboLigaseTMrequired to give 50% ligation of HindIII fragments of l DNA (5´ DNA termini concentration of 0.15 µM) in a total reaction volume of 20 µl in 30 minutes at 16°C in a buffer of 50 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 1 mM ATP, 10 mM dithiothreitol. 

TurboLigaseTM Specific Activity and Purity:
TurboLigase has a specific activity >12,000 unit/mg. TurboLigase is purified to homogeneity.

General
Component Formulation TurboLigase™: 5 unit/ul in Storage Buffer of 10 mM Tris-HCl, pH 7.5, 50 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, and 50% glycerol, 5X TurboLigation Buffer: 250 mM Tris-HCl, pH 7.5, 50 mM MgCl2, 5 mM dithiothreitol, 5 mM ATP, 25% polyethylene glycol 8000
Concentration 5 u/ul
Purity TurboLigase is purified to homogeneity.
Size 500 reactions
Specific Activity TurboLigase has a specific activity >12,000 unit/mg.
Storage and shelf-life TurboLigase and 5X TurboLigation Buffer are stable at -20°C for 12 months from date of receipt. Store TurboLigase and 5X TurboLigation Buffer at -20°C.
Unit Definition 0.015 Weiss unit (1 cohesive end ligation unit) is defined as the amount of TurboLigase™ required to give 50% ligation of HindIII fragments of I DNA (5' DNA termini concentration of 0.15 uM) in a total reaction volume of 20 ul in 30 minutes at 16°C in a buffer of 50 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 1 mM ATP, 10 mM dithiothreitol.
Source Species T4 phage

Ligation Reaction Protocol:

  • Warm 5X TurboLigation Buffer to room temperature (22o-25oC) before setting up TurboLigation reactions.
  • Keep TurboLigase at -20oC.
  • Thaw competent cells (not included) on ice immediately before setting up TurboLigation reactions.
  • Always include a control reaction with vector only.

 

Set up ligation reaction in     10 µl  

5X TurboLigation Buffer

vector

insert 

    2 µl 

__ µl    total vector and insert volume up to 7 µl

__ µl    use ~1:3 molar ratio of vector to insert

adjust volume with water to 

TurboLigase

 9 µl   

 1 µl    add TurboLigase last to increase efficiency

  

  • Mix gently and incubate at room temperature for 5 minutes. Incubation between 5 to 10 min gives the best result.
  • Chill the reaction on ice for 1 minute.
  • Use 1-3 µl of the ligation reaction for transformation of 20-50 µl competenet cells.

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