A molecular beacon probe1 has its natural fluorescence quenched in solution unless it is hybridized to the target sequence. Consequently, the design of a molecular beacon requires a fluorophore to be in one part of the sequence and the quencher molecule to be in another, with both molecules being separated from the oligonucleotide by a hydrocarbon spacer. The Dabcyl group has been found to be a universal quencher. 3'-Dabsyl CPG and 3'-Dabcyl CPG are used to prepare probes with the quencher blocking the 3'-terminus. 5'-Dabcyl Phosphoramidite locates the quencher at the 5'-terminus and Dabcyl-dT places it within the sequence, leaving the 3'-terminus available for polymerase extension.
Details
Usage
Coupling: Dissolve this product in anhydrous acetonitrile:THF (9:1). |A 6 minute coupling time is recommended.
Deprotection: No changes needed from standard method recommended by synthesizer manufacturer.
Specifications
Diluent
10% THF in Anhydrous Acetonitrile
Storage
Refrigerated storage, maximum of 2-8°C, dry
Stability
2-3 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
10-1058-02
0.25 g
.25grams
2.17
59
35.4
22.13
16.09
11.8
2.95
10-1058-90
100 µmol
.115grams
1
20
12
7.5
5.45
4
1
10-1058-95
50 µmol
.058grams
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
10-1058-02
0.25 g
.25grams
3.24
0.07
58.4
36.5
26.55
3.65
10-1058-90
100 µmol
.115grams
1.5
0.07
23.6
14.75
10.73
1.48
10-1058-95
50 µmol
.058grams
0.75
0.07
8.6
5.38
3.91
0.54
References
1 S. Tyagi and F.R. Kramer, Nature Biotechnology, 1996, 4, 303-308.
