Quasar® 670-dT Phosphoramidite - (10-5955)
5'-Dimethoxytrityloxy-5-[(N-(6-(N1-ethyl-3,3,3',3'-tetramethylindodicarbocyanin-1'-yl)hexanoyl)- aminohexyl)-3-acrylimido]-2'-deoxyUridine-3'-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite, hexafluorophosphate saltQuasar®570 and Quasar®670 dyes are two cyanine derivatives of Cyanine 3 and Cyanine 5, respectively. These two fluorophores differ in structure simply by the number of carbons in the conjugated polyene linkage. Cyanine dyes are a popular oligonucleotide modification and are commonly used for imaging, flow cytometry, and genomic applications. With Quasar®570-dT and Quasar®670-dT, these dyes can be incorporated internally without disrupting base pairing or stacking. Monomers can be inserted as a replacement for a dT residue.
Particularly for Quasar® 670-dT, the use of UltraMILD monomers is preferred. (Catalog Numbers: dA: 10-1601-xx, dC: 10-1015-xx, dG: 10-1621-xx, dT: 10-1030-xx). To avoid any exchange of the iPr-Pac group on the dG with acetyl, use the UltraMILD Cap Mix A (40-4210-xx/40-4212-xx). 0.02M Iodine for oxidation is recommended.
Usage
- Coupling: 10 minute coupling time recommended. The use of UltraMILD monomers are preferred. (Catalog Numbers: dA: 10-1601-xx, dC: 10-1015-xx, dG: 10-1621-xx, dT: 10-1030-xx). To avoid any exchange of the iPr-Pac group on the dG with acetyl, use the UltraMILD Cap Mix A (40-4210-xx/40-4212-xx). 0.02M Iodine for oxidation is recommended.
- Deprotection: If UltraMILD reagents were used, deprotect in 0.05M Potassium Carbonate in Methanol for 4 hours at Room Temperature OR for 2 hours at Room Temperature in 30% Ammonium Hydroxide. If standard bases were used, deprotection in Ammonium Hydroxide at Room Temperature for 24 hours will give acceptable yields. However, the oligonucleotide will require additional purification.
Specifications | |
---|---|
Diluent | Anhydrous Acetonitrile |
Recommended Storage | Freezer storage, -10 to -30°C, dry |
Stability In Solution | 1-2 Days |
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 | ||||
10-5955-95 | 50 µmol | 0.08 | 0.5 | 3.33 | 2 | 1.25 | 0.91 | 0.67 | 0.17 |
10-5955-90 | 100 µmol | 0.15 | 1 | 20 | 12 | 7.5 | 5.45 | 4 | 1 |
10-5955-02 | 0.25 g | 0.25 | 1.64 | 41.33 | 24.8 | 15.5 | 11.27 | 8.27 | 2.07 |
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