Hieff™ Versatile T4 DNA Ligase (High Fidelity, 600 U/µL)- 12996ES

YeasenSKU: 12996ES42

Size: 60 KU
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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
  • Thermal stable

 

Specifications

Source

Recombinant E.coli

Storage Buffer

50 mM Tris-HC1,100 mM KC1,0.1 mM EDTA,1 mM DTT,50%Glycerol,pH 7.50.2 @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

Component

 

Name

12996ES60 

60000 U

(60 KU)

12996ES62

600000U

(600 KU)

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.

Notes

1. Please operate with lab coats and disposable glovesfor 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

VolumeμL

Final concentration

dA-tailed DNA

60

10×T4 DNA Ligase Buffer

10

0.1 μM-0.5 μM

50% PEG6000

12*

1 ng -400 ng

DNA Adapter

5**

 

Hieff® Versatile T4 DNA Ligase (600 U/µL)

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

0.1 μM

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.

 

 

 

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