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JOE-dT Phosphoramidite - (10-5936)

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

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10-5936

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Product Specifications

Formula:

C₈₁H₉₁Cl₂N₆O₁₉P

M.W.:

1554.5

F.W.:

945.65

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

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
Recommended Storage	Freezer storage, -10 to -30°C, dry
Stability In Solution	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