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Epicentre® T4 RNA链接酶Ⅱ,突变体删除(T4 RNA Ligase 2, Deletion Mutant)上线

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Epicentre® T4 RNA链接酶Ⅱ,突变体删除(T4 RNA Ligase 2, Deletion Mutant)上线

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【摘要】:
T4RNALigase2,DeletionMutantApplicationsPreparingcDNAlibrariesforsmall-RNAtranscriptomeanalysissuchasRNA-Seq.OptimallinkerligationformiRNAcloning.T4RNALigase2,DeletionMutant,alsoknownasT4Rnl2(1-249),is

T4 RNA Ligase 2, Deletion Mutant

Applications

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

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 prevents circularization and other undesirable bimolecular reactions.

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.