What is SYBR Green II?
SYBR Green II RNA gel stain is a sensitive nucleic acid gel stain that has bright fluorescence when bound to RNA and low background in gels, making it ideal for use with either formaldehyde/agarose or polyacrylamide gels using laser scanners or standard UV transilluminators.
How does SYBR green stain DNA?
SYBR Green I binds to DNA. The resulting DNA-dye-complex best absorbs 497 nanometer blue light (λmax = 497 nm) and emits green light (λmax = 520 nm). The stain preferentially binds to double-stranded DNA, but will stain single-stranded (ss) DNA with lower performance.
How does SYBR Green work in qPCR?
SYBR Green is one of the most commonly used fluorescent dyes in qPCR. It binds to double-stranded DNA molecules by intercalating between the DNA bases. Once intercalated to DNA, SYBR Green becomes less mobile, causing its energy to be released as fluorescence.
How does SYBR Green qPCR work?
What wavelength is SYBR Green?
497 nm
The maximum excitation wavelength of SYBR Green I is 497 nm, but there is also a secondary excitation peak near 254 nm. The fluorescence emission of SYBR Green I stained DNA is centered at 520 nm. The dye is supplied as a 10,000× solution in dimethyl sulfoxide (DMSO).
What is the role of SYBR Green in qPCR?
SYBR® Green is a dsDNA-binding dye that intercalates nonspecifically into dsDNA, allowing measurement of the amount of PCR product. Since the increase in fluorescence is proportional to the amount of product accumulated, SYBR® Green qPCR can be used for relative DNA quantification.
Does SYBR Green inhibit PCR?
SYBR Green exhibits a very strong fluorescent signal, but it has been shown to inhibit the PCR reaction and has a narrow dynamic range and lower reproducibility than other detection chemistries [2, 3, 7].
What is SYBR Green assay?
SYBR Green I is the most commonly used fluorescent dye. It binds specifically to double-stranded DNA. Using this dye, double-stranded DNA molecules can be exclusively quantified in the presence of single-stranded DNA molecules during denaturation experiments.
How do you make SYBR Green?
Dilute the Sybr® Green I reagent 1:100 in 1X TE pH 8.0. (This 1:100 dilution has shown to be stable for up to 6 hours at room temperature when protected from light.) Example: For a 1:100 dye dilution transfer 10 ul of 10,000 X concentrated Sybr® Green I dye and 990 ul of 1X TE buffer into a clean amber or foiled1.
What kind of RNA is in SYBR Green 2 gel?
1% Agarose gel containing 16S and 23S ribosomal RNA (rRNA). SYBR® Green II RNA gel stain. Comparison of RNA detection in nondenaturing gels using SYBR® Green II RNA gel stain and ethidium bromide. DNA microarrays stained with nucleic acid stains for quality control. Supplied as a 10,000X concentrate in DMSO.
What are the spectral characteristics of SYBR Green I stain?
Spectral Characteristics SYBR® Green I stain is maximally excited at 497 nm, but also has secondary excitation peaks at ~290 nm and ~380 nm (Figure 2). The fluorescence emission of SYBR® Green I stain bound to DNA is centered at 520 nm.
Where is the fluorescence emission of SYBR Green I?
The fluorescence emission of SYBR® Green I stain bound to DNA is centered at 520 nm. These spectral characteristics make SYBR® Green I stain compatible with a wide variety of gel reading instruments, ranging from those with ultraviolet epi- and transillumination to argon laser and mercury-arc lamp excitation gel scanners.
Why is SYBR Green used in Bio-Rad?
SYBR ® Green has a high specificity for dsDNA and is especially useful when RNA or ssDNA may also be present in the sample. SYBR ® Green displays very low background fluorescence, and its excitation and emission spectra are well matched to the blue light or UV sources and filter sets in commonly used imaging instruments.