T4 DNA Ligase

Catalog# BPC1101-1
Lot # Check on the product label
Size: 100 U
Storage: Store at -20°C for 6 months. 
Concentration: 5 U/µl
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Price: $40.00

  


Catalog# BPC1101-1


Lot # Check on the product label


Size: 100 U


Storage: Store at -20°C for 6 months. 


Concentration: 5 U/µl


Application:

(1) Quick connection between DNA fragments and carrier DNA;

(2) Cloning of restriction enzyme cut fragments;

(3) Connect the DNA fragment with Linker or Adaptor.


Description: T4 DNA Ligase is isolated and purified from Escherichia coli expressing T4 DNA Ligase gene after induction. It catalyzes the phosphodiester binding of the 5'-phosphate and 3' hydroxyl ends of adjacent DNA strands. The enzyme catalyzes the ligation of blunt-ended or sticky-ended DNA and repairs single-stranded nicks in single-stranded DNA in double-stranded DNA, RNA, and DNA/RNA hybrids.


Enzyme storage buffer: 20mM Tris-HCl (pH7.5), 0.1 mM EDTA, 1mM DTT, 50 mM KCl, 50% glycerol.


10× Ligation Buffer: 400 mM Tris-HCl, 100 mM MgCl 2, 100 mM DTT, 5 mM ATP(PH7.8), other components.


  • Recommended ligation reaction system(Take a 10 μl reaction system as an example):

Component

Standard reaction tube

positive control tube

Vector

μl

μl

PCR product

n ul **

/

Control insert DNA fragment

/

1ul

T4 DNA Ligase (5U/ul)

25U

25U

10×Ligation Buffer

1ul

1ul

Add deionized water to final volume

10ul

10ul

1. PCR reaction cycle settings:


Sticky end ligation: 22°C reaction for 10 minutes (can be extended to 1 hour as needed);


Blunt-end ligation: 22°C for 2 hours or 16°C overnight.

Details