Description
Hieff™ Versatility T4 DNA Ligase is a variant of T4 DNA Ligase. This enzyme is suitable for catalyzing the ligation reaction between cohesive or blunt-ended molecules. It is a single-enzyme product designed for connecting DNA fragments and adapters during the library preparation process in next-generation sequencing(NGS). The Hieff® Versatility T4 DNA Ligase has been extensively validated through high-throughput sequencing and exhibits exceptional quality.
Feature
- High fidelity
- Thermotolerant
Learn more about the versatile T4 DNA ligase development
Specifications
Source | Recombinant E.coli |
Storage Buffer | 50 mM Tris-HC1,100 mM KC1,0.1 mM EDTA,1 mM DTT,50%Glycerol,pH 7.5士0.2 @25℃ |
Unit Definition | In a ligation reaction system of 20 μL, the enzyme amount required to catalyze the ligation of more than 50% of DNA fragments, when 6 μg of λDNA-Hind III is reacted at 16°C for 30 minutes, is defined as 1 unit of activity (U). |
Components
Components | Name |
12996ES60 60KU |
12996ES62 600KU |
12996-A | Hieff® Versatile T4 DNA Ligase (600 U/µL) | 100 μL | 1000 μL |
12996-B | 10×T4 DNA Ligase Buffer | 500 μL | 3×1500 μL |
Storage
This product should be stored at -25~-15℃ for 2 years.
Figure
Please leave a message to inquire about the performance comparison data between this enzyme and other top-tier thermostable and conventional T4 DNA ligases.
Notes
1. Please operate with lab coats and disposable gloves,for your safety.
2. If a small amount of precipitation occurs in the buffer, it is a normal phenomenon. Please invert and mix the buffer thoroughly before using it.
3. This product is for research use only.
Protocol
1. The current mainstream DNA library preparation kits using the ligation-based method typically do not require purification steps after end repair and A-tailing. The DNA fragments are directly subjected to adapter ligation without purification. This kit is compatible with the mainstream commercially available end repair and A-tailing systems for seamless adapter ligation.
Set up the following reaction in a microcentrifuge tube on ice, gently mix by pipetting or shaking, Centrifuge briefly and heat inactivate at 20°C for 15 minutes.
Table 1. Reaction System for Adapter Ligation
Component |
Volume(μL) |
Final concentration |
dA-tailed DNA |
60 |
1× |
10×T4 DNA Ligase Buffer |
10 |
0.1 μM-0.5 μM |
50% PEG6000 |
12* |
1 ng -400 ng |
DNA Adapter |
5** |
|
3~5*** |
|
|
Nuclease-free water |
up to 100 |
- |
【Note】*The 50% PEG6000 solution is not provided in the kit, you will need to prepare it yourself
**Please refer to Table 2 for the amount of adapter.
***The amount of T4 DNA Ligase can be added as needed by 3-5 μL.
2. The concentration of the adapter directly affects the ligation efficiency and library yield. Excessive use of adapters may result in more adapter dimers, while low usage may affect connection efficiency and library production. Table 2 lists the recommended adapter amount for different amounts of input DNA.
Table 2. The recommended adapter amount for 100 pg-1 μg Input DNA
Input DNA |
Concentration |
0.1 ng |
|
1 ng |
0.5 μM |
10 ng |
1 μM |
25 ng |
2 μM |
100 ng~1000 ng |
10 μM |
【Note】:The amount of adpter can be adjusted according to the requirements in the table above.