SIMA (dichloro-diphenyl-fluorescein) is a xanthene dye with spectral characteristics similar to those of HEX but with a higher quantum yield. SIMA has higher stability during deprotection under alkaline conditions, so deprotection can be run with aqueous ammonium hydroxide at higher temperatures or with AMA (1:1 mixture, concentrated aqueous ammonium hydroxide/40% aqueous methylamine) at room temperature for 2 h or 65°C for 10 min. When used for deprotection with aqueous ammonium hydroxide at 55°C overnight, oligonucleotide-bound SIMA does not degrade, while with HEX the fluorophore degrades by at least 10%.
SIMA phosphoramidite is used in oligonucleotide synthesis to produce fluorescently labeled primers and hybridization probes for quantitative PCR.
Spectral properties
Excitation/absorption maximum, nm
531
ε, L⋅mol⁻¹⋅cm⁻¹
92300
Emission maximum, nm
555
Fluorescence quantum yield
0.63
CF₂₆₀
0.57
CF₂₈₀
0.18
Oligo synthesis details
Diluent
anhydrous acetonitrile
Coupling conditions
coupling time 3 min
Deprotection conditions
Standard conditions with 25% ammonium hydroxide; deprotection time depends on the composition of nucleic acids and their protective groups. AMA (1:1 mixture of concentrated aqueous ammonium hydroxide / 40% aqueous methylamine) can be used for 2 hours at room temperature or 10 min at 65°C.
规格
规格或纯度
BioReagent, 用于显微镜, 适用于荧光分析, 生物染色剂, ≥95%(HPLC)
稳定性与储存
Store at -20℃ long term (12 months). Store in the dark. Desiccated.
危险品化学品经营许可证（带存储）