SYBR Green I Nucleic Acid Gel Stain 

MCE SYBR Green I Nucleic Acid Gel Stain is a proprietary unsymmetrical cyanine dye that has proven exceptionally useful for assays requiring sensitive nucleic acid detection. SYBR Green I stain is maximally excited at 490 nm and has secondary excitation peaks at 290 nm and 380 nm. The fluorescence emission of SYBR Green I Stain bound to DNA is centered at 520 nm. The detection limit using SYBR Green I Stain is as low as 60 pg per band of dsDNA using 300 nm transillumination. With 254 nm epi-illumination, as little as 20 pg of dsDNA can be detected. SYBR Green I Stain is also possible to stain dsDNA prior to electrophoresis as it has an exceptionally high affinity for dsDNA.

•   High sensitivity: Detect as little as 20 pg dsDNA per band

•   High signal-to-noise ratio: strong fluorescence signal of the sample, low background signal

•   Strong applicability: suitable for agarose gel or polyacrylamide gel electrophoresis; used for dsDNA, ssDNA or RNA staining

•   No destaining or washing steps, compatible with different testing equipments

Stored at -20°C, and is stable for 1 year.

Protect from light.

Staining DNA Before Electrophoresis

a. Precast agarose or nondenaturing polyacrylamide gels (0.8-3.0%) with SYBR Green I stain by diluting the SYBR Green I stock reagent 1:10,000 into the gel solution just prior to pouring the gel.

b. Mix SYBR Green I stain with 6× Loading Buffer at a ratio of 1:9 to get Loading Buffer pre-dye. Then mix the nucleic acid sample and marker with Loading Buffer pre-dye at a ratio of 5:1 respectively and incubate at room temperature for at least 15 minutes prior to electrophoresis.

Note: Loading Buffer containing SDS would affect the electrophoresis results. It is recommended to use a Loading Buffer without SDS.

c. After loading and electrophoresis, detect the bands under UV illuminator.

Staining DNA Following Electrophoresis

a. Perform electrophoresis on an agarose or nondenaturing polyacrylamide gel without stain.

b. Dilute the stock SYBR Green I reagent with a buffer of pH 7.5-8.0 (such as TAE, TBE or TE) at a certain ratio (the ratio between dye and agarose gel is 1:10,000 or 2:10,000; the ratio between dye and polyacrylamide gel is 3:10,000 or 4:10,000) and gently shake to form a staining solution.

c. Cover the gel with staining solution in a plastic container and incubate at room temperature for 30-60 minutes with shaking. Protect the staining container from light by covering it with aluminum foil or placing it in the dark. Staining time varies depending on the thickness of the gel and the percentage of agarose or polyacrylamide.

d. Detect the bands under UV illuminator.