SYBR Safe™ versus ethidium bromide

1. SYBR Safe™ versus ethidium bromide Nikita Warner, Ph.D. Assoc. Product Manager Nucleic Acid Separation & Analysis

2. A DNA gel stain that is: Better for you displays no acute oral toxicity and greatly reduced genotocixity Better for the environment offers non-hazardous waste status and Clean Water Act compliance Better for your institution reduces waste storage and disposal costs Better for your sample improves cloning efficiency when used with blue-light illumination without compromising detection sensitivity as compared to ethidium bromide What is SYBR Safe™? SYBR Safe™ is a better DNA gel stain

3. How is SYBR Safe™ better for me?

4. Oral toxicity evaluation Displays no acute oral toxicity

5. Genotoxicity studies Displays greatly reduced genotocixity

6. Ames test results Causes fewer mutations in the standard Ames test

7. How is SYBR Safe™ better for the environment?

8. Hazardous waste testing • According to the guidelines of the EPA, SYBR Safe™ DNA gel stain is not • corrosive • ignitable, or • reactive Not classified as hazardous waste under U.S. federal regulations

9. NPDES analysis Tests performed according to National Pollutant Discharge Elimination System (NPDES) guidelines indicate that SYBR Safe™ DNA gel stain does not differ significantly from 0.5X TBE buffer SYBR Safe™ DNA gel stain does not differ significantly from 0.5X TBE buffer

10. Organic compound analysis • Tested were performed on 0.5X TBE and 1X SYBR Safe™ in 0.5X TBE for: • Organochlorine pesticides and PCBs (608M) • Volatile organic compounds (624) • Semi-volatile organic compounds (625) None of the organic pollutant compounds tested for were detected in either sample SYBR Safe™ DNA gel stain in 0.5X TBE is indistinguishable from 0.5X TBE alone

11. How is SYBR Safe™ better for my institution?

12. SYBR Safe™ Institutional Savings Case Study: Forsyth Institute, Boston, MA (data provided by Forsyth Institute) For more details, visit: www.invitrogen.com/sybrsafe 14% institutional savings at a Boston institute

13. SYBR Safe™ Waste Disposal at ISU Iowa State University, Ames IA • Wastewater officials in Ames, Iowa approved the disposal of SYBR Safe™ as nontoxic waste • Diluted dye solution can be poured directly down the drain at Iowa State University ISU scientists can dispose SYBR Safe™ liquid waste down the drain

14. How is SYBR Safe™ better for my sample?

15. Sensitivity comparison Kb 23.1 9.4 6.6 4.4 2.3 2.0 Ethidium bromide SYBR Safe™ Marker is l DNA/Hind III Fragment SYBR Safe™ has comparable sensitivity to ethidium bromide

16. E-Gel® - EtBr versus SYBR Safe™ 1.2% E-Gel® with ethidium bromide 0.3 second exposure Same samples and volumes loaded on both gels. Both gels viewed on UV transilluminator 1.2% E-Gel® with SYBR Safe™ 1 second exposure Same sensitivity as ethidium bromide, but longer exposure

17. How does one use SYBR Safe™?

18. Using SYBR Safe™ • Formats offered: • Offered as 10,000X concentrate, or • Ready-to-use solution in 1X TAE • Ready-to-use solution in 0.5X TBE • E-Gel® pre-cast agarose gels incorporated with SYBR Safe™ • How to use: • Incorporate into the gel, or • Perform post-run staining, or • Use E-Gel® with SYBR Safe™ SYBR Safe™ is convenient to use

19. What does UV exposure do to DNA?

20. Nicked DNA Supercoil DNA Effect of UV on DNA SYBR SafeTM & Blue Light Ethidium bromide & UV Light 2 4 8 16 Time (Min) 2 4 8 16 CFUs obtained from transformation with constructs containing inserts stained with SYBR Safe™ or ethidium bromide and viewed on blue light or UV respectively UV damages DNA and diminishes cloning results

21. Eliminate the dangers of UV transillumination Safe Imager Blue light Transilluminator • Does not damage your skin and eyes • Produces bright light and uniform emission • Provides optimal excitation for SYBR Safe™ DNA gel stain • For more information, visit www.invitrogen.com/safeimager Optimum excitation of SYBR Safe™ and maximum safety

22. Safe Imager versus the competition Competitor blue light box Safe Imager™ technology Even illumination with the Safe Imager™

23. 0.25 500 256 128 .125 0.5 16 32 64 2 1 4 8 Safe Imager for visualizing stained proteins SYPRO Ruby staining ng protein per band (approx.) 2x dilution series Mark 12 Markers SafeImager 1 ng sensitivity with SYPRO Ruby and SafeImager

24. 500 128 256 0.5 16 32 0.125 1 2 4 8 0.25 64 Safe Imager and SYPRO-Ruby Staining of 2-D gel Mark12 standards Jurkat cell lysate Avoid UV exposure when performing 2-D electrophoresis

25. Safe Imager and SYPRO Orange protein stain 128 256 128 256 0.5 0.5 16 32 32 16 1 0.125 1 2 0.125 4 2 0.25 8 0.25 8 4 64 64 SYPRO Orange (Safe Imager) Lucy 565 (UV) Superior sensitivity with SYPRO Orange and Safe Imager versus Sigma’s Lucy 565

26. 128 256 0.5 32 16 0.125 1 2 4 0.25 8 64 256 128 0.5 32 16 0.125 1 2 4 8 0.25 64 Safe Imager and SYPRO Ruby protein stain Lucy 506 (Safe Imager) SYPRO Ruby (Safe Imager) Mark12 standards Superior sensitivity with SYPRO Ruby and Safe Imager versus Sigma’s Lucy 506

27. The complete safety solution SYBR Safe™ DNA Gel Stain • Better for you • Better for the environment • Better for your institution • Better for your sample • All this with … • No compromise on performance • For more information, visit • www.invitrogen.com/sybrsafe Safe Imager Blue light Transilluminator • Avoid UV exposure • Get improved cloning efficiency • Can be used with DNA and protein stains SYBR Safe™ and Safe Imager™ offer the compete safety solution

28. Information Learn more about: SYBR Safe™ at www.invitrogen.com/sybrsafe Safe Imager™ at www.invitrogen.com/safeimager Additional information or questions: E-mail sybrsafe@invitrogen.com We like you just the way you are!