[{"group": "General", "name": "Materials Required but not Supplied RNase Inhibitor, Murine (NEB #M0314) (optional) Nuclease-free Water (NEB #B1500) 100% DMSO Unit Definition One unit is defined as the amount of enzyme required to convert 1 nanomole of 5´- [ 32 P rA 16 into a phosphatase-resistant form in 30 minutes at 37°C Reaction Conditions 1X T4 RNA Ligase Reaction Buffer Supplement with 1 mM ATP 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 50 mM KCl 10 mM Tris-HCl 0.1 mM EDTA 1 mM DTT 50% Glycerol pH 7.5 @ 25°C Heat Inactivation 65°C for 15 minutes Unit Assay Conditions 1X T4 RNA Ligase reaction buffer, supplemented with 1 mM ATP, is mixed with the RNA substrate (10μM of 5´-[ 32 P]rA 16 ) and varying amounts of enzyme. Incubation is at 37°C for 15 minutes (8).", "value": ""}]
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