Accelagen
 
작성일 : 12-05-07
[Accelagen] TurboLigation Kit
 
TurboLigationTM Kit
 
A. Produc detail
 
1. Description
TurboLigationTM Kit enables quick ligation of both blunt-end or cohesive-end of DNA fragments in only 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 minutes reaction is as efficient as a traditional 16℃ overnight ligation reaction. The reaction product is used directly for chemical transformation without any further treatment.
 
2. Applications
    - Adding linkers or adapters to DNA
    - Blunt or cohesive end ligation of duplex DNA
    - DNA cloning
 
3. 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
 
4. Stability and Storage
TurboLigase and 5X TurboLigation Buffer are stable at -20℃ for 12 month from date of receipt.
Store TurboLigase and 5X TurboLigation Buffer at -20℃.
Shipped in gel packs.
 
 
B. Data sheet
 
1. TurboLigaseTM Unit Definition
0.015 Weiss unit (1 cohesive end ligation unit) is defined as the amount of TurboLigaseTM required 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.
 
2. TurboLigaseTM Specific Activity and Purity
TurboLigase has a specific activity >12,000 unit/mg.
TurboLigase is purified to homogeneity.
 
 
 
C. Protocol
 
1. Ligation Reaction
- Warm 5X TurboLigation Buffer to room temperature (22℃-25℃) before setting up TurboLigation reactions.
- Keep TurboLigase at -20℃.
- 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       2 µl
vector                                  µl              total vector and insert volume up to 7 µl
insert                                   µl              use ~1:3 molar ratio of vector to insert
                                                                                                                                                                
adjust volume with water to 9 µl
TurboLigase                       1 µl               adding TurboLigase the last increases ligation 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.
 
 
Ordering information
 
Catalog No.
Product Name
Size / (Conc.)
 L0110S
TurboLigationTM Kit
100 reactions
L0110M
500 reactions
L0110B
Bulk
 
 
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