Recommendations to reduce SYBR® Green I (4250-050-05) fading
Posted: Tue 17 Feb 2015 12:48Are there recommendations to reduce SYBR® Green I (4250-050-05) fading?
FAQ from Trevigen
1. Apply diluted SYBR® Green I solution and dry in the dark before viewing.
The stain is more stable and still proportional to DNA in the sample.
2. Apply additional SYBR® Green I solution without washing off first application.
3. Avoid prolonged exposure to fluorescent light from microscope.
4. Prepare Anti-fade Solution, as described, and apply 10 µl per sample, covering samples with coverslip.
In a 50 ml tube, mix until dissolved:
p-Phenylenediamine dihydrochloride 500 mg
1X PBS 4.5 ml
Add approximately 400 µl of 10 N NaOH dropwise with stirring until pH of solution reaches 7.5-8.0. Add 1X PBS to increase the volume to 5 ml, and 45 ml of glycerol for a final volume of 50 ml.
Vortex mixture thoroughly and apply 10 µl per sample, covering samples with coverslip.
Nail polish may be used to seal coverslip. Re-staining of slides is not recommended.
Store anti-fade solution at -20ºC for up to one month. Darkening of solution may occur.
5. Add beta-mercaptoethanol (1 mM) to diluted dye.
FAQ from Trevigen
1. Apply diluted SYBR® Green I solution and dry in the dark before viewing.
The stain is more stable and still proportional to DNA in the sample.
2. Apply additional SYBR® Green I solution without washing off first application.
3. Avoid prolonged exposure to fluorescent light from microscope.
4. Prepare Anti-fade Solution, as described, and apply 10 µl per sample, covering samples with coverslip.
In a 50 ml tube, mix until dissolved:
p-Phenylenediamine dihydrochloride 500 mg
1X PBS 4.5 ml
Add approximately 400 µl of 10 N NaOH dropwise with stirring until pH of solution reaches 7.5-8.0. Add 1X PBS to increase the volume to 5 ml, and 45 ml of glycerol for a final volume of 50 ml.
Vortex mixture thoroughly and apply 10 µl per sample, covering samples with coverslip.
Nail polish may be used to seal coverslip. Re-staining of slides is not recommended.
Store anti-fade solution at -20ºC for up to one month. Darkening of solution may occur.
5. Add beta-mercaptoethanol (1 mM) to diluted dye.