Quasar® 670-dT Phosphoramidite
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 salt
Technical Documents
Safety Data Sheet
Glen Report 33-25: New Products — Dye-dT Phosphoramidites
Description
Quasar®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.
Details
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.
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 |
Expedite
| Catalog # | Pack Size | Grams/Pack | Dilution (mL) | Approximate Number of Additions |
|---|
| Molarity | 50nm | 0.2μm | 1μm | 15μm |
|---|
| 10-5955-95E | 50 µmol | 0.08 | 0.75 | 0.07 | 8.6 | 5.38 | 3.91 | 0.54 |
| 10-5955-90E | 100 µmol | 0.15 | 1.5 | 0.07 | 23.6 | 14.75 | 10.73 | 1.48 |
| 10-5955-02E | 0.25 g | 0.25 | 2.45 | 0.07 | 42.6 | 26.63 | 19.36 | 2.66 |
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