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Gelred and Gelgreen GelRed and GelGreen Safe and highly sensitive nucleic acid gel stains for replacing toxic EtBr Ethidium Bromide (EtBr) −gel stain of the past Problems: • Highly toxic (mutagenic)! H2N NH2 • Expensive to dispose + N Br- • High background signal (destaining may be necessary) • Limited sensitivity GelRed and GelGreen Are the Perfect Replacements for EtBr Five Simple Reasons 1. Safety: Nonmutagenic and noncytotoxic 2. Easy disposal: Safe to dispose in the drain 3. Compatibility: Spectrally compatible with existing instruments 4. Sensitivity: Higher signal but lower background 5. Stability: can be stored at RT and microwavable Reason #1. Safety • GelRed and GelGreen are structurally engineered to minimize chances of interacting with genomic DNA in live cells GelRed and GelGreen are impenetrable to latex gloves Glove Penetration Test: 48 hr dialysis against 5X GelRed or GelGreen 1x (ref.) 140 6000 1X (ref.) 120 5000 ce ence 90) 100 4000 sc 5 538) e escen r r m o o Em 80 E u u 3000 l l 4/ F 54 e x 485/ 60 ve F x v i E i t 2000 ( E t a ( l a l e dialyzed e 40 R R dialyzed 1000 20 0 0 Ge lGre en GelRe d Conclusion: handling GelRed or GelGreen with latex glove is safe. GelRed and GelGreen are impenetrable to cell membranes HeLa cells were stained with 1X SYBR Safe, GelRed or GelGreen in PBS for 30 minutes. Images were taken on an Olympus mercury arc lamp microscope using appropriate filters. SYBR Safe GelRed GelGreen Bright Field view Fluorescence Field view GelRed and GelGreen Are Nonmutagenic − Largely a consequence of cell membrane impermeability 1200 EtBr GelGreen GelRed 1000 s Comparison of mutagenicity among GelRed, e 800 GelGreen and EtBr. Ames tests performed in oni l Salmonella strain TA98 with S9 metabolic o C 600 activation of r e b m 400 Concentration range used in gel staining u N 200 *** 0 0 0.1 0.25 0.5 1 2.5 5 7.5 10 25 50 Dose (ug/plate) * EB not tested at these concentrations (More Ames test results in GelRed/GelGreen Safety Report) Reason #2. Easy Disposal • GelRed and GelGreen successfully passed the Aquatic Toxicity Test (CCR Title 22) based on the EPA/600/4-85/013 protocol; thus, they can be disposed safely down the drain (See safety report for detailed results) Reason #3. Instrument Compatibility • GelRed and EtBr have similar absorption spectra 1.8 1.6 EtBr 1.4 GelRed 1.2 1 s Ab 0.8 0.6 0.4 0.2 0 220 320 420 520 620 Wavelength (nm) Conclusion: Just like EtBr, GelRed can be perfectly excited by an UV transilluminator. GelRed and EtBr Have Nearly Identical Emission Spectra EtBr(EM), GelRed(EM) 400 e 300 nc ce es 200 r o u l F 100 0 nm 400 500 600 700 800 Wavelength (nm) Conclusion: Replacing EtBr with GelRed requires no filter change. GelGreen Is A Visible Light-excitable And Green Fluorescent Gel Stain n tio n o i rp s o is Abs Em 250 350 450 550 650 Wavelength (nm) Conclusion: 1) Although GelGreen can be excited by UV light, it is ideal for laser-based gel scanners or dark reader using visible light excitation; 2) Visible light excitation reduces DNA damage Reason #4. Superior Sensitivity Comparison of GelRed and EtBr in Post Gel Staining GelRed EtBr High signal, Limited signal, Low background High background Comparison of GelRed and EtBr in Precast Gel Staining GelRed EtBr High background in high Mwt. DNA region High signal, Low background Insensitive to small DNA fragments, Comparison of GelGreen and SYBR Safe GelGreen SYBR Safe Conclusion: GelGreen is far more sensitive than SYBR Safe Reason #5. Both GelRed and GelGreen Have Superior Stability • Stable in water indefinitely • Can be stored at room temperature • Can be microwaved or heated in agarose buffers • GelRed is more photostable than EtBr • GelGreen is more photostable than SYBR Safe Summary • GelRed is a direct and the best replacement for EtBr • GelGreen is ideal for gel staining using visible light excitation and requiring post DNA recovery.
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