[{"group": "General", "name": "Unit Definition 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 (#S1315) in a total reaction volume of 20 μl in 1 hour at 25°C. 5´-FAM-rArGrUrCrGrUrArGrCrCrUrUrUrArUrCrCrGrArGrArUrUrCrArGrCrArArUrA-3´ 5´-rAppCTGTAGGCACCATCAAT–NH2-3´ Molarity = 14 μM Reaction Conditions 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) Storage Buffer 10 mM Tris-HCl 100 mM NaCl 1 mM DTT 0.1 mM EDTA 50% Glycerol pH 7.5 @ 25°C Heat Inactivation 65°C for 20 minutes Unit Assay Conditions 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.", "value": ""}]
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