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ROX Phos-CPG Column (5'-DMT-mdC(TEG-ROX)-Phos-CPG)

ROX Phos-CPG Column (5'-DMT-mdC(TEG-ROX)-Phos-CPG)

CPG column used to incorporate a methyl-dC ROX-phosphate moiety into the 3' end of an oligonucleotide.
  • Used to add a passive, stable reference dye to the 3' end of an oligonucleotide.
  • Excitation/emission maxima of 586 nm and 610 nm, comparable with the SYBR Green I or TaqMan probes, making it ideal to be used with our BHQ-2 quencher.
  • 500 Å CPG suitable for high yield applications such as therapeutic oligos (≤ 30mers).
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Product information

ROX, a member of the rhodamine family, is routinely used as a passive reference dye to provide a stable baseline for the fluorescent reporter dyes, i.e SYBR Green I or TaqMan probes, in multiplex qPCR or real-time PCR. The baseline also allows the correction of pipetting errors, fluorescence fluctuations, and instrumental drift such as change of lamp intensity output over time. As the dye does not interfere qPCR amplification, a constant amount is added to all samples as a control. Dependant on the filter and real-time PCR instrument used during experiments, concentration of ROX may need be to be adjusted. Earlier real-time PCR instruments did not have filter matching ROX and thus high concentrations were needed to establish the baseline. With the ROX dye being compatible with all PCR instruments, the later instruments are now calibrated for ROX spectral settings and thus removes the need to re-calibrate the thermal cycler. With an excitation/emission maxima of 586 and 610, the dye has a different emission spectrum compared against the SYBR Green I or TaqMan probes, making it ideal to be used with our BHQ-2 quencher. Compared to the fluorescein family of dyes, rhodamine dyes are more resistant to photo bleaching and stable over a wider pH range.

Properties:

  • Appearance: purple powder
  • Absorption Maximum (Lambda Max): 575
  • Extinction Coefficient at Lambda max: 91000
  • Extinction Coefficient at 260 nm: 38500
  • Fluorescence Maximum: 602

Product usage:

  • Synthesis conditions: Follow the instrument manufacturer's protocol, with noted changes, when using this product. Synthesis can be performed on these supports according to standard procedures with deprotection and cleavage modifications noted.
  • Cleavage conditions: Use t-butylamine/water (1:3) for 45 minutes at 25 °C. The use of ammonia must be avoided, as it will cause degradation of the ROX moiety.
  • Deprotection conditions: When using fast deprotecting amidites (eg. C-Ac, G-DMF, G-PAC) use t-butylamine/Water (1:3) for 6 hours at 65° C. When using standard amidites (eg. C-Bz, G-iBu) use t-butylamine/Water (1:3) overnight (12-15 hours) at 65°C. The use of ammonia must be avoided, as it will cause degradation of the ROX moiety.
  • Image of cleaved and deprotected structure:
  • The mass this product adds after conjugation and work-up (the additional mass seen by mass spectrometry) is: 1103.06

Storage and handling:

  • Shipping conditions: Ambient
  • Storage conditions: -15 to -30 °C in sealed container

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