6-TAMRA SE → 6-Carboxytetramethylrhodamine

HRID 871792Notes

Reaction equation

HRID 871792 reaction equation
[CH3:1][N:2]([C:4]1[CH:9]=[CH:8][C:7]2[C:10]([C:21]3[CH:26]=[C:25]([C:27]([O:29]N4C(=O)CCC4=O)=[O:28])[CH:24]=[CH:23][C:22]=3[C:37]([O-:39])=[O:38])=[C:11]3[C:19]([O:20][C:6]=2[CH:5]=1)=[CH:18][C:14](=[N+:15]([CH3:17])[CH3:16])[CH:13]=[CH:12]3)[CH3:3].[O-]P(OP(OP([O-])([O-])=O)([O-])=O)(=O)[O-]>CS(C)=O.C([O-])([O-])=O.[Na+].[Na+].C([O-])(O)=O.[Na+]>[CH3:17][N:15]([C:14]1[CH:13]=[CH:12][C:11]2[C:10]([C:21]3[CH:26]=[C:25]([C:27]([O-:29])=[O:28])[CH:24]=[CH:23][C:22]=3[C:37]([OH:39])=[O:38])=[C:7]3[C:6]([O:20][C:19]=2[CH:18]=1)=[CH:5][C:4](=[N+:2]([CH3:1])[CH3:3])[CH:9]=[CH:8]3)[CH3:16]

Participants

5

Reactant 2

HCID 27626066-TAMRA SEHCID 3440921Triphosphate

Product 1

HCID 27626046-Carboxytetramethylrhodamine

Reagents, catalysts & solvents 2

HCID 679Dimethyl SulfoxideSolventHCID 10791Sodium SesquicarbonateSolvent

Procedure

A solution of 6-TAMRA-SE (0.75 mg, 1.4 μmol) in anhydrous DMSO (30 μL) was added to a solution of triphosphate dC.10 (1.6 μmol) in Na2CO3/NaHCO3 buffer (0.1 M, pH 9.2; 0.3 mL) and incubated at room temperature for 30 minutes. The reaction was purified with reverse-phase HPLC using a Perkin Elmer OD-300 C18 column (4.6×250 mm) to yield the 6-TAMRA labeled triphosphate WW2p080. Mobile phase: A, 100 mM triethylammonium acetate (TEAA) in water (pH 7.0); B, 100 mM TEAA in water/CH3CN (30:70). HPLC purification was achieved a linear gradient of 5-50% B for 40 minutes and then 50-90% B for 10 minutes. The concentration of WW2p080 was estimated by adsorption spectroscopy using the extinction coefficient of the 6-TAMRA dye (i.e., 65,000 at 555 nm).

Workup

  1. customThe reaction was purified with reverse-phase

Sources & evidence

ORD record
ord-5efb8781c4b143339c252fa23c2aa42c
Source dataset
uspto-grants-2015_03
Patent number
US08969535B2

Reaction notes

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