NEB, M0351S, T4 RNA Ligase 2, truncated K227Q
T4 RNA Ligase 2, truncated K227Q ligates the pre-adenylated 5´ end of DNA or RNA to the 3´ end of RNA.
Catalog Number: M0351S
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Category
Pack Size200 units / pack
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| <b>Unit Definition</b> 200 units is defined as the amount of enzyme required to give 80% ligation of a 31-mer RNA to the pre-adenylated end of a 17-mer DNA Universal miRNA Cloning Linker ( NEB #S1315 ) in a total reaction volume of 20 μl in 1 hour at 25°C. 5´-FAM- rArGrUrCrGrUrArGrCrCrUrUrUrArUrCrCrGrArGrArUrUrCrArGrCrArArUrA-3´ 5´-rAppCTGTAGGCACCATCAAT–NH 2 -3´ Molarity | 14 µM Concentration: 200,000 units/ml <b>Reaction Conditions</b> 1X T4 RNA Ligase Reaction Buffer Incubate at 25°C 1X T4 RNA Ligase Reaction Buffer 50 mM Tris-HCl 10 mM MgCl2 1 mM DTT (pH 7.5 @ 25°C) <b>Storage Buffer</b> 10 mM Tris-HCl 100 mM NaCl 1 mM DTT 0.1 mM EDTA 50% Glycerol pH 7.5 @ 25°C <b>Heat Inactivation</b> 65°C for 20 minutes <b>Molecular Weight</b> Theoretical: 71406.97 daltons <b>Specific Activity</b> 200,000 units/mg <b>Unit Assay Conditions</b> 1X T4 RNA Ligase Reaction Buffer supplemented to 10% (w/v) PEG MW 8000, 20 pmol of 5´-FAM labeled RNA, and 40 pmol preadenylated DNA linker. After incubation at 25°C for 1 hour, the ligated product is detected on a 15% denaturing polyacrylamide gel. |
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