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SYTOX™ Green Nucleic Acid Stain - 5 mM Solution in DMSO - FAQs

View additional product information for SYTOX™ Green Nucleic Acid Stain - 5 mM Solution in DMSO - FAQs (S7020)

3 product FAQs found

How do SYTO dyes bind to DNA?

The binding mode of SYTO nucleic acid stains is unknown. However, the behavior of these and related nucleic acid dyes suggests the following binding properties:

1.They appear to contact the solvent (suggested by sensitivity to salt, divalent cations, and in particular, SDS) and thus are likely to have contacts in the grooves.
2.All SYTO dyes appear to show some base selectivity and are thus likely to have minor groove contacts.
3.They can be removed from nucleic acid via ethanol precipitation; this characteristic is not shared by ethidium bromide and other intercalators. Likewise, the dyes are not removed from nucleic acid via butanol or chloroform extraction. These extraction methods do remove ethidium bromide from nucleic acid. 4. SYTO binding is not affected by nonionic detergents.
5. SYTO dyes are not quenched by BrdU, so they do not bind nucleic acids in precisely the same way as Hoechst 33342 and DAPI ((4′,6-diamidino-2-phenylindole).

SYBR Green I has shown little mutagenicity on frameshift indicator strains, indicating that it isn't likely to strongly intercalate.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I need to mount my Qdot secondary-labeled tissue samples in HistoMount mounting medium, but my Qdot 705 conjugate overlaps with your Qnuclear Deep Red nuclear label. What other nuclear label would you recommend, that is compatible with HistoMount mounting medium?

We recommend using SYTOX Green stain, which you can image using a FITC filter and we have shown to be compatible with HistoMount mounting medium. Some have used DAPI, but there have been some issues with slight quenching and spectral shifting of DAPI into green or even red wavelengths.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I am using SYTOX AAdvanced as a dead cell stain, but all of my cells are labeling even though I am certain that they are supposed to be alive. These are adherent cells that I have trypsinized. Why am I getting false-dead signals?

SYTOX AAdvanced labels only dead cells because it is a cell impermeant dye. The dye can only enter cells that have a compromised plasma membrane. Trypsinization may cause temporary disruption of the plasma membrane, sufficient to allow staining with a cell impermeant dye. You can reduce the “false-dead” problem by either reducing the amount of trypsin and/or reduce the incubation time for trypsinization or use a gentler dissociation reagent such as TrypLE Express, TrypLESelect reagents, or Versene. After trypsinization, wash well, and if possible, allow a recovery time in normal culture media before staining with any of the SYTOX dyes.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.