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
Safety Data Sheet
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 |
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