SYBR™ Green Nucleic Acid Gel Stain Starter Kit - FAQs

The SYBR dyes are useful only over a narrow range of pH, from about 7 to 8. Outside this range, the fluorescent signal diminishes rapidly.

Please go here (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/nucleic-acid-gel-electrophoresis/dna-stains/sybr-safe.html) and click on the “Filter Recommendations” tab to see filter recommendations for use with SYBR Safe DNA Gel Stain. Note that the excitation and emission spectra of SYBR Safe DNA Gel Stain are very similar to those of SYBR Green I, SYBR Green II, and SYBR Gold dyes, as well as fluorescein (FITC). Therefore, filters appropriate for these dyes can also be used. A camera filter is not required with the Safe Imager Blue-Light Transilluminator; the amber filter provided with the instrument serves this purpose.

This is not recommended. Most deep amber/orange ethidium bromide filters have a cutoff value around 550 nm. The SYBR Green dyes emit at 520 nm, which will not be detected using this filter.

All SYBR dyes have similar spectral properties, but have different chemical compositions. All SYBR dyes bind to dsDNA, ssDNA and RNA but vary in sensitivity. SYBR Safe DNA Gel Stain was specifically developed as a safer alternative to ethidium bromide. SYBR Green I is an ultrasensitive stain for dsDNA, and SYBR Green II is a highly sensitive stain for RNA and ssDNA. All SYBR dyes are optimally excited by the Safe Imager Blue-Light Transilluminator.

Disposal regulations vary. Please contact your safety office or local municipality for disposal guidelines.

Different SYBR dyes bind to dsDNA, ssDNA, and RNA, but vary in the sensitivity and specificity with which they bind to different nucleic acids. SYBR Green I Nucleic Acid Gel Stain is used for staining dsDNA and ssDNA. SYBR Green II RNA Gel Stain will stain dsDNA and ssDNA but has better sensitivity for RNA. SYBR Gold Acid Gel Stain was developed after SYBR Green I and II and is the most sensitive fluorescent gel stain offering the highest sensitivity for both DNA and RNA. SYBR Safe DNA Gel Stain is a reduced mutagenicity formula designed for use with blue light systems. It is less sensitive than the SYBR Green I and II but comparable to ethidium bromide.

Please check the following link for additional details regarding the different SYBR dyes: The Molecular Probes Handbook: Nucleic Acid Detection on Gels, Blots and Arrays

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

Yes, you can stain your TBE gels with ethidium bromide, SYBR Green I, SYBR Green II, and the SilverXpress Silver Staining Kit. For ethidium bromide staining, soak the gel in a 2 µg/mL solution of ethidium bromide in ultrapure water for 20 minutes. Destain by rinsing with three successive 10-minute rinses of ultrapure water. Visualize bands under UV light.