How do you stain SYBR green gel?
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. Can I use the ethidium bromide filters on my camera to image SYBR dyes?
Why is SYBR Green used in gel electrophoresis?
It is used as a dye for the quantification of double stranded DNA in some methods of quantitative PCR. It is also used to visualise DNA in gel electrophoresis. Higher concentrations of SYBR Green can be used to stain agarose gels in order to visualise the DNA present.
What is the use of SYBR Safe DNA gel stain?
SYBR safe DNA gel stain is a highly sensitive stain for visualization of dna in agarose or acrylamide gels. SYBR safe stain is specifically formulated to be a less hazardous alternative to ethidium bromide that can utilize either blue light or uv excitation.
Why is SYBR green better than ETBR?
SYBR® Safe It is marketed as being less harmful than ethidium bromide, but this is debatable. Its major advantage is that it is as sensitive as ethidium bromide but does not require UV light for visualization.
How does SYBR Green qPCR work?
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.
Is SYBR Green sensitive to light?
SYBRGreen is light sensitive, exposure to light should be minimize in case you still have some time before running.
Is SYBR Green toxic?
In numerous tests carried out by independent, licensed testing laboratories, SYBR Safe DNA Gel Stain showed little or no genotoxicity and no acute toxicity.
What does SYBR Green bind to?
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 much SYBR gel is safe?
Dilute SYBR Safe DNA Gel Stain concentrate 10,000-fold in TAE or TBE buffer prior to use. For most minigels, 50 mL of 1X stain is sufficient (e.g., dilute 5 µL of concentrate with 50 mL buffer). For larger gels, increase volumes proportionally, ensuring that the entire gel is fully immersed during staining.
How do you read SYBR Safe DNA gel stain?
DNA stained with SYBR Safe DNA gel stain can be viewed using a blue light transilluminator such as Invitrogen’s Safe Imager instrument, or a standard UV transilluminator. If you plan to use the DNA for cloning, avoid exposing DNA stained with SYBR Safe DNA gel stain to UV light.
Is SYBR Green Safe?
SYBR Safe DNA Gel Stain was specifically developed as a safer alternative to ethidium bromide. Invitrogen SYBR Green I Nucleic Acid Gel Stain is an ultrasensitive stain for dsDNA, and Invitrogen SYBR Green II RNA Gel Stain is a highly sensitive stain for RNA and ssDNA.
Is SYBR Green safer than ethidium bromide?
SYBR Safe. SYBR safe is a commercial DNA stain manufactured by Invitrogen. It is marketed as less harmful than ethidium bromide, but this is debatable. Its major advantage is that it is as sensitive as ethidium bromide but does not require UV light for visualization.
How long does SYBR Green i nucleic acid gel stain last?
SYBR® Green I nucleic acid gel stain (S7563) 500 µL 10,000X ≤–20°C Desiccate Protect from light • • • When stored as directed in DMSO, stain is stable for 6 months to 1 year.
Is the SYBR Safe gel stain visible to the eye?
Bands stained with SYBR Safe DNA Gel Stain are visible to the eye on a 300 nm transilluminator. However, optimum detection is obtained by photographing the gel using a UV-compatible emission filter with your CCD or film camera.
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.
What is the pH of SYBR Green I reagent?
Stain may be diluted in TE (10 mM Tris-HCl, 1 mM EDTA, pH 8.0), TBE, or TAE buffer. Staining with SYBR® Green I reagent is pH sensitive. For optimal sensitivity, verify that the pH of the staining solution at the temperature used for staining is between 7.5 and 8.0 (preferably pH 8.0).