JOE-dT Phosphoramidite

5'-Dimethoxytrityl-5-[6-(4',5'-Dichloro-2',7'-dimethoxy-3',6'-dipivaloylfluorescein-6-carboxamido)-hexanoyl)-3-aminoallyl]-2'-deoxyUridine-3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite

Product Specifications

Formula:
C81H91Cl2N6O19P
M.W.:
1554.50
F.W.:
945.65
To Retrieve a Catalog Number, Select a Pack Size and Format:

Technical Documents


Description

Phosphoramidites containing derivatives of fluorescein are designed to take advantage of the multicolor detection capability of modern DNA sequencers and genetic analyzers. Dichloro-dimethoxy-fluorescein (JOE) exhibits unique spectroscopic characteristics when compared to other derivates, with absorbance and emission maxima at 525 nm and 548 nm, respectively. JOE-dT can be inserted into the desired sequence as a replacement for a dT residue. Internal fluorophores offer a convenient method for introducing dyes in a sequence without disrupting normal base pairing.

Details

Usage

  • Coupling: 10 minute coupling time recommended.
  • Deprotection: Use Ammonium Hydroxide and deprotect as required by nucleobases. If AMA is used, a small amount of a non-fluorescent impurity will be formed. To eliminate this impurity, first deprotect with ammonium hydroxide for 30 minutes at room temperature, add an equal volume of 40% methylamine and then complete the deprotection as required by the nucleobases - e.g. 10 minutes at 65°C or 2 hours at room temperature for standard bases. Extended deprotection in ammonium hydroxide for 17 h at 55 °C also yields acceptable results.
Specifications
Diluent Anhydrous Acetonitrile
Storage Freezer storage, -10 to -30°C, dry
Stability 1-2 Days

Dilution/Coupling Data

The table below show pack size data and, for solutions, dilution and approximate coupling based on normal priming procedures.

ABI 392/394

Catalog # Pack Size Grams/Pack 0.1M Dil. (mL) Approximate Number of Additions
LV40 LV200 40nm 0.2μm 1μm 10μm
10-5936-02 0.25 g 0.25 1.61 40.33 24.2 15.13 11 8.07 2.02
10-5936-90 100 µmol 0.16 1 20 12 7.5 5.45 4 1
10-5936-95 50 µmol 0.08 0.5 3.33 2 1.25 0.91 0.67 0.17

Expedite

Catalog # Pack Size Grams/Pack Dilution (mL) Approximate Number of Additions
Molarity 50nm 0.2μm 1μm 15μm
10-5936-02E 0.25 g 0.25 2.4 0.07 41.6 26 18.91 2.6
10-5936-90E 100 µmol 0.16 1.5 0.07 23.6 14.75 10.73 1.48
10-5936-95E 50 µmol 0.08 0.75 0.07 8.6 5.38 3.91 0.54