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Enzymes for Molecular Biology

Enzymes for Molecular Biology: Enzymes for Molecular Biology

T4 RNA Ligase 2, Deletion Mutant

T4 RNA Ligase 2, Deletion Mutant, also known as T4Rnl2(1-249)

BioSearch Tech (Lucigen/Epicentre)

Catalogue No.DescriptionPack SizePriceQty
LR2D11310K T4 RNA Ligase 2, Deletion Mutant10kU £217.00 Quantity Add to Order
LR2D1132KT4 RNA Ligase 2, Deletion Mutant2kU £63.00 Quantity Add to Order

T4 RNA Ligase 2, Deletion Mutant

T4 RNA Ligase 2, Deletion Mutant, also known as T4Rnl2(1-249)

BioSearch Tech (Lucigen/Epicentre)

Ligate single-stranded adenylated DNA or RNA oligos to RNA targets for a variety of applications.

  • Ligate single-stranded adenylated DNA or RNA oligos to RNA targets
  • Use this enzyme to prepare cDNA libraries for small-RNA transcriptome analysis such as RNA-Seq
  • Perform linker ligation for miRNA cloning applications

T4 RNA Ligase 2, Deletion Mutant, also known as T4Rnl2(1-249), is used to ligate single-stranded adenylated DNA or RNA oligonucleotides to small RNAs for cloning or next-generation RNA sequencing. The preadenylated 5´ ends of DNA or RNA are ligated to the 3´ ends of RNA. Unlike the full-length enzyme, T4Rnl2(1-249) is unable to adenylate the 5´ end of the substrate in the presence of ATP. However, it can use a preactivated donor (App-DNA or App-RNA) and join it to the 3´ end of an acceptor; thus, performing the ligation reaction in the absence of ATP which prevents circularization and other undesirable bimolecular reactions.

The enzyme is available in 2,000 and 10,000-Unit sizes at a concentration of 200 U/µl. The enzyme is supplied with a 10X Reaction Buffer.

Unit Definition: One unit is the amount of enzyme required to give 50% ligation of a 22-mer RNA to the preadenylated end of a 17-mer DNA when both oligos are annealed to a complementary 39-mer DNA strand in 30 minutes at 37°C under standard assay conditions.

Storage Buffer: 50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1 mM EDTA, 1 mM dithiothreitol (DTT), and 0.1% Triton® X-100.

T4 RNA Ligase 2, Deletion Mutant, 10X Reaction Buffer: 500 mM Tris-HCl (pH 7.5), 20 mM MgCl2, and 10 mM DTT.

Activity Assay: The unit definition assay is performed in a reaction containing 50 mM Tris-HCl (pH 7.5), 2 mM MgCl2, 1 mM DTT and 0.4 µg of equimolar mix of 22-mer, 17-mer and 39-mer oligonucleotides, and varying amounts of enzyme.

Quality Control: T4 RNA Ligase 2, Deletion Mutant, is free of detectable DNA exo- and endonuclease, and RNase activities.

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

T4 RNA Ligase 2, Deletion Mutant

T4 RNA Ligase 2, Deletion Mutant, also known as T4Rnl2(1-249)

BioSearch Tech (Lucigen/Epicentre)

PROTOCOL

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

T4 RNA Ligase 2, Deletion Mutant

T4 RNA Ligase 2, Deletion Mutant, also known as T4Rnl2(1-249)

BioSearch Tech (Lucigen/Epicentre)

Applications

  • Preparing cDNA libraries for small-RNA transcriptome analysis such as RNA-Seq.
  • Optimal linker ligation for miRNA cloning.

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200