MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity Ethidium dsDNA 290 nm agarose 0.2ng-0.5ng May be UV Mutagenic with Managed Approx. $30 Bromide 1 ssDNA 605 nm stored at Transilluminator S9 activation as for 10mL of RNA acrylamide room of Salmonella hazardous 10mg/mL temperature; Polaroid 667 TA98 and waste solution; use PCR indefinite black & white TA1537 0.5µg/mL for storage strains agarose gel; yields 5,000 LD 50 , rat (oral): 40-mL gels, 1503 mg/kg $0.006/gel. (slightly toxic) Aquatic toxicity : LC 50 not available, MSDS indicates “may cause long- term adverse effects on aquatic environment” SYBR Safe 2 dsDNA 280/502 nm agarose Comparable Keep away UV Weakly Approved $53.75 for ssDNA 530 nm to ethidium from heat Transilluminator mutagenic with by MWRA 10,000X acrylamide bromide and light; S9 activation for drain SYBR Safe stable for blue (vis) light of Salmonella disposal, in DMSO, pre-cast approx. 6 transilluminator TA98 and May 2005 400µL; 4µL months (SafeImager™ TA1537 for 40 mL post-gel when stored recommended) strains gel = at room $0.5375 per temperature. laser scanner LD 50 , rat (oral): gel. >5,000 mg/kg Polaroid 667 (practically black & white nontoxic) 1 Available from numerous vendors 2 Invitrogen/Molecular Probes (now Life Technologies) QUESTIONS??? Send comments to [email protected] 1 MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity with SYBR Safe filter (S37100) or Aquatic Molecular toxicity : Probes SYPRO LC 50 >750mg/L filter (S6656) or Kodak Wratten #9 filter GelRed 3 dsDNA 275 nm agarose 1-3ug/mL; Store in UV Weakly Approved Available as ssDNA 600 nm visualizes water at Transilluminator mutagenic at by MWRA 0.5mL RNA acrylamide DNA doses room 18.5 µg/mL for drain (500µL) of as low as temperature similar spectra with S9 disposal, 10,000X pre-cast 25ng. or at 4 oC as ethidium activation of April 2010 concentrate; (agarose) bromide, Salmonella working therefore can TA98 dilution is 1- post-gel use same 3µg/mL for (both) visualization LD 50 , rat (oral): 167-500 settings not provided minigels on MSDS $100-$105 Cell ($0.20- membrane $0.63/gel) staining: did not cross HeLa cell membranes after 30 minutes incubation Aquatic toxicity : LC 50 >750mg/L GelGreen 3 dsDNA 250- agarose 1-3µg/mL Store in UV Non- Approved 500µL ssDNA 300/500nm water at Transilluminator mutagenic by MWRA 10,000X RNA ~525 nm acrylamide room (254 nm) from 0.4µg/mL for drain concentrate 3 Biotium QUESTIONS??? Send comments to [email protected] 2 MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity – not temperature to 18.5 µg/mL disposal, in DMSO (GelRed recommended or at 4 oC 488nm laser- in Salmonella April 2010 recommended due to slow based gel TA98 and 1-3µg/mL for for ssDNA and diffusion of scanner TA1537 167-500 RNA) dye strains, w/ and minigels w/o S9 pre-cast activation. $90-$100 ($0.18- post-gel LD 50 , rat (oral): $0.60/gel); not provided on MSDS Cell membrane staining: did not cross HeLa cell membranes after 30 minutes incubation Aquatic toxicity : LC 50 >750mg/L MegaFluor 4 dsDNA Similar agarose Can run at Store 10X UV Manufacturer Not $225 for absorption 3pg-5pg gel at 4 oC transilluminator claims present in 500-1500 -added directly to and post-gel (picograms) molecule is the gel gels = DNA sample, not emission (agarose) similar spectra large and does running $0.15-$0.45 gel or buffer spectra as Can as ethidium not cross cell buffer; per gel ethidium visualize bromide, membrane. buffer -not advised for bromide DNA doses therefore can Indicated may be Dose : denatured DNA less than use same testing disposed MegaFluor 10ng if visualization performed on by drain. to DNA, 1:10 samples are settings epithelial cells 4 GenTaur QUESTIONS??? Send comments to [email protected] 3 MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity first showed no No formal MegaFluor incubated at mutagencity. MWRA to DNA 60 oC. approval; marker, 2:10 LD 50 , rat( oral) : meets or not stated exceeds MegaFluor criteria for to genomic Aquatic SYBR DNA, 4:10 toxicity : Safe. LC 50 not stated 1.5 mL = 1,500 dsDNA gels @10µL (100 samples per 1.5mL kit) SafeWhite 5 dsDNA 320 nm agarose 0.1ng-0.3ng Store 6X in Same as for Non- Approved 6X in water; ssDNA 480 nm water at 4 oC ethidium bromide mutagenic in by MWRA 1mL, $58 RNA Salmonella for drain TA98 and disposal, Dose : 5µL TA1537 April 2010 per 100 mL strains, w/ and agarose = w/o S9 $0.29 per activation. gel LD 50 , rat( oral) : not stated Aquatic toxicity : LC 50 not stated SafeRed 4 dsDNA 490 nm agarose 0.3ng-0.8ng Store 6X in Same as for Non- Approved 6X in water; ssDNA 630 nm water at 4 oC ethidium bromide mutagenic in by MWRA 1 mL, $58 RNA Salmonella for drain TA98 and disposal, Dose : 5µL TA1537 April 2010 per 100 mL 5 Applied Biological Materials QUESTIONS??? Send comments to [email protected] 4 MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity strains, w/ and agarose = w/o S9 $0.29 per activation. gel LD 50 , mouse subcutaneous : 16mg/kg Aquatic toxicity : LC 50 not stated SafeGreen 4 dsDNA 290 nm agarose 0.2ng-0.6ng Store 6X in Same as for Non- Approved 6X in water; ssDNA 490 nm water at 4 oC ethidium bromide mutagenic in by MWRA 1 mL, $58 RNA Salmonella for drain TA98 and disposal, Dose : 5µL TA1537 April per 100 mL strains, w/ and 2010. agarose = w/o S9 $0.29 per activation. gel LD 50 , mouse subcutaneous : 250 mg/kg Aquatic toxicity : LC 50 not stated GelStar 6 dsDNA 493 nm agarose 20pg for Lasts 12 302nm or 312 Mutagenicity Treat as 10,000X in ssDNA 527 nm dsDNA; months nm filter for UV testing data potential DMSO, RNA acrylamide 25pg for when stored transilluminator not provided mutagen $158 for ssDNA; at -20 oC. per 2x250µL pre-cast 10pg for GelStar filter for LD 50 , rat( oral) : direction (agarose) RNA. photographs not stated of product Dose : DNA, maker. 4µL per 6 Cambrex QUESTIONS??? Send comments to [email protected] 5 MIT EHS Office Green Chemistry/Pollution Prevention Program Ethidium Bromide Alternatives Assessment August 2009 (revised: August 2011) Product Nucleic acids Visual Gel base and Sensitivity Stability or Types of visual Mutagenicity, Disposal Unit price, visualized and range, nm application (ug/mL) or Storage equipment Acute dose & cost method as (i.e., precast lowest Limits (gel Toxicity and per gel absorb/emit or post-gel) dilution documentation) Aquatic (abs/em) reported Toxicity post-gel Aquatic 40mL gel = toxicity : 125 LC 50 not stated gels@$1.27 per gel RNA, 8µL per 40mL gel = 62.5 gels@ $2.53 per gel. QUESTIONS??? Send comments to [email protected] 6 .