'Safe’ is the latest addition to the SYBR® stain family and has a number of very attractive benefits:
Increased safety - Data released by Molecular Probes show that SYBR Safe stain is negative in 3 different mammalian cell-based assays for genotoxicity and exhibits low mutagenicity compared to EtBr in Ames tests. Furthermore, SYBR safe has been extensively tested for environmental safety and is NOT classified as hazardous waste under U.S. Federal regulations.
Flexible usage - SYBR Safe can be used either to stain a gel after running or, for the fastest results, it can be used as an‘ in-agarose’ stain.
High stability - Safe is stable for over 6 months in aqueous solution and is also stable to the heating involved in preparing molten agarose.
Adding SYBR Safe to the agarose before electrophoresis has a number of major advantages:
- DNA bands can be directly visualized as they migrate through the gel using the Dark Reader GelHead. An electrophoresis run can be halted as soon as the desired DNA bands are separated - often within 30 minutes.
- Alternatively, if using a Dark Reader transilluminator, the bands can be visualized immediately after the electrophoresis run without any wait for staining to occur.
- Because SYBR Safe is stable to both the heating involved in preparing molten agarose, as well as storage in aqueous solution, a batch of agarose containing stain can be prepared ahead of time and stored in aliquots for immediate use whenever needed. This feature makes SYBR Safe the quickest and most convenient of all the SYBR dyes.
0.5 g of agarose was added directly to 50 mL of SYBR Safe stain solution and heated in a microwave for about 1 minute. The 1% agarose solution was then poured into the bed of a Dark Reader GelHead.
The gel was loaded with a 2-fold dilution series of a 1 kb DNA ladder (ISC BioExpress) starting with 100 ng of total DNA and then run at 200 V for 30 minutes using 0.5 x TBE running buffer.
The DNA detection sensitivity using SYBR Safe is comparable to EtBr/UV - about 1 ng by eye and 0.5 ng using a CCD camera.
