T4 DNA Ligase for NGS

Item Number

T665501

• Storage Temp: Store at -20°C,Avoid repeated freezing and thawing
• Shipped In: Ice chest + Ice pads

Product Description

T4 DNA Ligase is isolated and purified from Escherichia coli expressing the T4 DNA Ligase gene after induction and expression. It catalyzes the binding reaction of adjacent DNA strands with 5 'phosphate groups and 3' hydroxyl groups via phosphodiester bonds. This enzyme can catalyze the connection of flat or sticky end DNA, repairing single stranded incisions in double stranded DNA, RNA, and DNA/RNA hybridization, but has no activity for single stranded nucleotides.

T665501	Component	1500 U	7500 U	Storage
T665501A	T4 DNA Ligase, 15 U/μL	100 μL	500 μL	-20°C. Avoid freeze/thaw cycle.
T665501B	4×T4 DNA Ligase Buffer	600 μL	2×1.5 mL	-20°C. Avoid freeze/thaw cycle.

Activity definition:

1U refers to the amount of enzyme required to convert 1nmol [32PPi] into Norit absorbable form within 20 minutes at 37 ° C in the ATP-PPi exchange reaction, equivalent to approximately 200 viscous terminal junction units.

Application scope:

Mainly used for adapter connection during the construction process of NGS Chinese library.

Usage:

It is recommended to use Kangwei Adaptor for connection, or choose to use Adaptors from NEB or Illumina companies. For specific connection methods, please refer to the product manuals of each company. The following are the steps to connect using Kangwei Adaptor:
1. Directly add the following reagents to the reaction solution that has completed DNA end repair:

Reagents Name	volume
4×T4 DNA ligase Buffer	25 μL
T4 DNA ligase ,15 U/μL	5 μL
Adaptor	5 μL
ddH2 O	Supplemented to 50 μ L

At this point, the total volume of solution in the tube is 100 μ L.
Note: If the initial sample size is less than 100 ng, please dilute the adapter 10 times with deionized water to 1.5 μ Use after M.
2. Use a gun to blow and aspirate the above reagents, mix well, and centrifuge briefly to collect the solution to the bottom of the tube.
Take a warm bath at 

3.23 ℃ for 20 minutes.
Attention: If using a PCR instrument for this operation, please close the heat cap.
4. Continue with subsequent steps, such as selective recovery of DNA fragments or purification of DNA fragments.