Immobilized T4 DNA Ligase
NEBU 65 Heat

  • Immobilized T4 DNA Ligase (IM T4 DNA Ligase) consists of T4 DNA Ligase covalently bound to magnetic beads. 
  • Easily remove enzyme from reaction using a magnet 
  • Avoid heat inactivation step, which may interfere with downstream applications
  • Re-use immobilized enzymes 
  • Enable new workflows and applications
Catalog # Concentration Size
M0569S 10 mg/ml 1.1 mg
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  • Product Information

    T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5´ phosphate and 3´ hydroxyl termini in duplex DNA or RNA. T4 DNA Ligase joins blunt end and cohesive end termini as well as repairs single stranded nicks in duplex DNA and some DNA/RNA hybrids (1).

    Immobilized T4 DNA Ligase (IM T4 DNA Ligase) is a slurry of magnetic beads coated with T4 DNA Ligase to produce a 10 mg/ml solution (50% glycerol) with an effective concentration of 60 cohesive end units (CEU) per microliter of slurry. Following a reaction, enzyme can be removed using a magnet, and can be re-used.

     

    Relative Activity of Immobilized T4 DNA Ligase at different temperatures

    Temperature

    % Activity

    16ºC

    100%

    25ºC

    100%

    37ºC

    50%


    Activity of Immobilized T4 DNA Ligase in Different NEB Buffers

    Buffer

    pH

    % Activity

    NEBuffer r1.1

    7.0

    50%

    NEBuffer r 2.1

    7.9

    100%

    NEBuffer r3.1

    7.9

    70%

    rCutSmart Buffer

    7.9

    100%

    Phi29 DNA Polymerase Buffer

    7.5

    100%

    Standard Taq Buffer

    8.3

    70%

    Thermopol Buffer

    8.8

    70%

    Q5 Reaction Buffer

    8.5

    10%

    T4 DNA Ligase Buffer

    7.5

    100%

    NEBNext Quick Ligation Buffer

    7.6

    25%

     


    Figure 1: Successful ligation of Lambda DNA- HindIII Digest by Immobilized T4 DNA Ligase



    Ligations of Lambda DNA- HindIII Digest (NEB #N3012) were performed with either no ligase, soluble T4 DNA Ligase (NEB #M0202) or Immobilized T4 DNA Ligase (NEB #M0569) according to recommended protocols.  Results indicate ligation is equivalent between soluble and immobilized enzyme.



    Figure 2:  Immobilized T4 DNA Ligase can be re-used 



    Consecutive splint ligations of Nanopore Adaptors were performed with Immobilized T4 DNA Ligase (NEB #M0569) in Quick Ligation Buffer. The data shows that activity remains unaffected after 20 consecutive ligation reactions. 

    Reagents Supplied

    The following reagents are supplied with this product:

    NEB # Component Name Component # Stored at (┬░C) Amount Concentration
    Product Categories:
    DNA Ligases Products,
    DNA Modifying Enzymes & Cloning Technologies Products
    Applications:
    Non-Cloning Ligation,
    DNA Ligation,
    Cloning Ligation,
    Cloning & Synthetic Biology
    • Properties & Usage
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