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NEB/Immobilized T4 DNA Ligase/1.1 mg/M0569S
  • NEB/Immobilized T4 DNA Ligase/1.1 mg/M0569S

NEB/Immobilized T4 DNA Ligase/1.1 mg/M0569S

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货号: M0569S
品牌: NEB
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  • 使用说明
  • 常见问题
    • 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 LigaseBuffer

      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 unaffectedafter 20 consecutive ligation reactions.
      This product is related to the following categories:
      DNA Ligases Products
      This product can be used in the following applications:
      Non-Cloning Ligation,
      DNA Ligation,
      Cloning Ligation
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