Spermidine → Digoxigenin
Reaction equation
[Mg+2].[Cl-].[Cl-].S[CH2:5][C@@H:6]([C@@H:8]([CH2:10]S)O)[OH:7].N[CH2:13][CH2:14][CH2:15][CH2:16]NCCCN.[Na+].[Cl-].P(OC[C@H:38]1[O:42][C@@H:41](N2C3N=CN=C(N)C=3N=C2)[C@H:40](O)[C@@H:39]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.P(O[CH2:68][C@H:69]1[O:73][C@@H:72](N2C3N=C(N)NC(=O)C=3N=C2)[C@H:71](O)[C@@H:70]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.C1C(=O)NC(=O)N([C@@H:95]2O[C@H:98](COP(OP(OP(O)(O)=O)(O)=O)(O)=O)[C@@H:97](O)[C@H:96]2O)C=1.C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)[CH2:119][C:120](O)=[O:121].C(O)C(N)(CO)C[OH:139].Cl>C(O)C.O>[CH3:95][C@@:96]12[C@H:10]3[CH2:119][C@@H:120]([OH:121])[C@:14]4([CH3:13])[C@@H:15]([C:39]5[CH2:38][O:42][C:41](=[O:139])[CH:40]=5)[CH2:16][CH2:5][C@:6]4([OH:7])[C@@H:8]3[CH2:68][CH2:69][C@@H:70]1[CH2:71][C@@H:72]([OH:73])[CH2:98][CH2:97]2Participants
Reactant 9
Product 1
Reagents, catalysts & solvents 2
Procedure
Transcription reactions were performed using the DIG RNA Labeling Kit (Roche Molecular Biochemicals, Indianapolis, Ind.). Transcription reactions consisted of 300 ng of purified PCR product, 10×Transcription Buffer (400 mM Tris-HCl pH 8.0; 60 mM MgCl2, 100 mM dithioerythritol, 20 mM spermidine, 100 mM NaCl, 1 unit/ml RNase inhibitor), DIG Labeling mix (10 mM each of ATP, CTP, GTP, and 6.5 mM UTP, 3.5 mM UTP-DIG; in Tris-HCl pH 7.5), T7 or T3 polymerase (20 units/μl) (Promega, Madison, Wis.), and molecular grade water. Reactions were carried out at 37° C. for 2 hours, stopped with 0.2 M EDTA, pH 8.0, ethanol precipitated, and resuspended in molecular grade water to a final concentration of approximately 1 μg/μl and stored at −20° C. until use.
Workup
- customprecipitated
- customstored at −20° C. until use
Sources & evidence
- ORD record
- ord-d98d4556161848b7af824b4d1cf23bb9
- Source dataset
- uspto-grants-2008_08
- Patent number
- US07416848B2
Reaction notes
Loading…