Summary of Health and Safety Test Results - Invitrogen

Summary of Health and Safety Test Results
SYBR Safe DNA Gel Stain

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Mens Sana in Corpore Sano.
(translation: A healthy mind in a healthy body.)
SYBR Safe DNA gel stain promotes safety by removing a
known health hazard—ethidium bromide—from your lab.
Ethidium bromide is so commonplace in molecular biology labs
that it’s easy to forget how hazardous it is. However, ethidium
bromide is a known carcinogen and a powerful mutagen,
making it both a health hazard and a disposal headache. There
has long been a need for a safer nucleic acid gel stain, and now
it’s here. Specifi cally developed to enhance lab safety without
compromising staining performance, SYBR Safe DNA gel stain
is less mutagenic, less toxic, more environmentally friendly, and
it works with traditional gel staining methods.
Further Information
SYBR Safe DNA gel stain provides:
n Safety—Less mutagenic than ethidium bromide;
tests negative for genotoxicity and acute oral toxicity
(LD 50 >5000 mg/kg)
n Low Environmental Impact—Not classifi ed as a hazardous
waste; meets the requirements of the Clean Water Act
n Sensitivity—Uncompromised detection sensitivity as com-
pared to ethidium bromide
n Convenience—Provided as a concentrate or as a ready-to-use
solution in TBE or TAE buff er; can be precast in gels or used as
a post-stain
n Speed—No destaining required; can be cast directly in the gel
n Compatibility—Use with a standard UV epi- or trans-
illuminator, a visible-light transilluminator, or a laser-based
scanner
This document is intended to present a brief summary of the safety and environmental data
gathered so far about our SYBR Safe DNA gel stain. Further information—including detailed
performance and safety results—can be accessed at probes.invitrogen.com/sybrsafe.
We want to help you solve your disposal problems.
If your local disposal agencies require additional
tests or data, please let us know by sending an
e-mail to sybrsafe@invitrogen.com.
Important Licensing Information
These products may be covered by one or more Limited Use Label Licenses (see the Invitrogen website,
www.invitrogen.com, or the Molecular Probes website, probes.invitrogen.com). By use of these products
you accept the terms and conditions of all applicable Limited Use Label Licenses. All products are for
research use only. CAUTION: Not intended for human or animal diagnostic or therapeutic uses.

Feeling Lucky?
Overview of SYBR Safe Test Results
SYBR Safe DNA gel stain does not induce transformations in
primary cultures of Syrian hamster embryo (SHE) cells when
compared with solvent alone, strongly indicating that the SYBR
Safe stain is noncarcinogenic. In contrast, ethidium bromide
tests positive in the SHE assay, consistent with its known activity
as a strong mutagen.
SYBR Safe DNA gel stain does not cause mutations in
mouse lymphoma cells at the TK locus, nor does it induce
chromosomal aberrations in cultured human peripheral blood
lymphocytes, with or without S9 metabolic activation, using
standardized tests against appropriate controls.
Important Licensing Information
These products may be covered by one or more Limited Use Label Licenses (see the Invitrogen website,
www.invitrogen.com, or the Molecular Probes website, probes.invitrogen.com). By use of these products
you accept the terms and conditions of all applicable Limited Use Label Licenses. All products are for
research use only. CAUTION: Not intended for human or animal diagnostic or therapeutic uses.
SYBR Safe DNA Gel Stain
Compared to ethidium bromide, SYBR Safe DNA gel stain
causes fewer mutations in the standard Ames test, as mea-
sured in several diff erent strains of Salmonella typhimurium.
Weakly positive results for SYBR Safe DNA gel stain in this test
occurred in four out of seven strains and only with activation by
a mammalian S9 fraction.
A single oral administration of SYBR Safe DNA gel stain
produces no signs of mortality or toxicity at a limit dose of
5000 mg/kg.
Based on extensive environmental safety testing, SYBR Safe
DNA gel stain is not classifi ed as hazardous waste under U.S.
Federal regulations (Resource Conservation and Recovery Act
(RCRA)). SYBR Safe DNA gel stain meets the requirements
of the Clean Water Act and the National Pollutant Discharge
Elimination System (NPDES) requirements.
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Born to Be Wild-Type.
Genotoxicity Results
Conclusion
SYBR Safe DNA gel stain does not induce transformations in
primary cultures of Syrian hamster embryo (SHE) cells when
compared with solvent alone, strongly indicating that the SYBR
Safe stain is noncarcinogenic. In contrast, ethidium bromide
tests positive in the SHE assay, consistent with its known activity
as a strong mutagen.
Summary of Results
An independent laboratory investigated the ability of
SYBR Safe DNA stain for inducing an increase in morpho-
logical transformation of Syrian hamster embryo (SHE) cells,
relative to vehicle control cultures, following a 7-day exposure
period. None of the three treatment groups (0.0500, 0.150, and
0.300 μg/ml) induced a signifi cant increase in morphological
transformation compared to the concurrent vehicle control. In
addition, a signifi cant increase of the morphological transfor-
mation frequency was also obtained from the positive control
treatment with benzo α pyrene at 5.0 mg/ml. The test article
(SYBR Safe stain) was therefore evaluated as negative in the
screening SHE cell transformation assay under 7-day exposure
conditions of this study.
Table 1. SYBR Safe stain—Mammalian genotoxicity analysis
Conclusion
SYBR Safe DNA gel stain does not cause mutations in mouse
lymphoma cells at the TK locus, nor does it induce chro-
mosomal aberrations in cultured human peripheral blood
lymphocytes, with or without S9 metabolic activation, using
standardized tests against appropriate controls.
Summary of Results
An independent laboratory investigated the ability of
SYBR Safe DNA gel stain to induce chromosomal aberrations
in cultured peripheral blood lymphocytes with and without
exogenous metabolic activation. No signifi cant increases in
the number of cells with structural aberrations, polyploidy, or
endoreduplication were observed in test either with or without
S9 activation. Similarly, no increases in the mutant frequency
were observed that exceeded the minimum criteria in L5178Y
TK+/- mouse lymphoma cells.
In In In Vitro Vitro Vitro Test * Cell Type Result with S9 Activation † Result without S9 Activation †
Transformation 1 Syrian hamster embryo (SHE) cells NA Negative
Chromosomal aberrations 2 Cultured human peripheral blood lymphocytes Negative Negative
Forward mutation 3,4 L5178YTK+/- mouse lymphoma cells Negative Negative
* In vitro tests were performed by Covance Laboratories Inc., Vienna, VA. † Mammalian S9 fraction obtained from Aroclor 1254–induced rat liver.
NA = Not applicable.
1. Fundamental and Molecular Mechanisms of Mutagenesis 356, 1 (1996); 2. Evans, H.J., in Chemical Mutagens, Principles and Methods for Their Detection,
A. Hollaender, Ed., Vol. 4, (1976) pp. 1–29; 3. Mutation Res 72, 447 (1980); 4. Mutation Res 59, 61 (1979).
Important Licensing Information
These products may be covered by one or more Limited Use Label Licenses (see the Invitrogen website,
www.invitrogen.com, or the Molecular Probes website, probes.invitrogen.com). By use of these products
you accept the terms and conditions of all applicable Limited Use Label Licenses. All products are for
research use only. CAUTION: Not intended for human or animal diagnostic or therapeutic uses.

Conclusion
Compared to ethidium bromide, SYBR Safe DNA gel stain
causes fewer mutations in the standard Ames test, as mea-
sured in several diff erent strains of Salmonella typhimurium.
Weakly positive results for SYBR Safe DNA gel stain in this test
occurred in four out of seven strains and only with activation by
a mammalian S9 fraction (see fi gure).
Summary of Results
Samples were pretreated with a mammalian S9 fraction and
then tested. With S. S. S. typhimurium typhimurium typhimurium strains TA97a, TA98, TA100
and TA102, an increase in revertants of more than twofold over
background indicates a positive result for mutagenicity in this
test. With strains TA1535, TA1537 and TA1538, an increase in
revertants of more than threefold over background indicates a
positive result.
We Like You Just the Way You Are.
Toxicity Results
Conclusion
A single oral administration of SYBR Safe DNA gel stain produces
no signs of mortality or toxicity at a limit dose of 5000 mg/kg.
Table 2. SYBR Safe stain—Toxicity analysis
Analysis Method Results
Aquatic toxicity * Fathead minnow
CA Title 22 acute screening
Oral toxicity ** EPA Acute Oral Toxicity Test
OPPTS 870.1100
Not hazardous or toxic
to aquatic life
LD50 > 5000 mg/kg
* Performed by AMEC Earth and Environmental San Diego Bioassay Laboratory, San Diego, CA.
** Peformed by Northview Pacifi c Laboratories, Inc., Hercules, CA.
Important Licensing Information
These products may be covered by one or more Limited Use Label Licenses (see the Invitrogen website,
www.invitrogen.com, or the Molecular Probes website, probes.invitrogen.com). By use of these products
you accept the terms and conditions of all applicable Limited Use Label Licenses. All products are for
research use only. CAUTION: Not intended for human or animal diagnostic or therapeutic uses.
Fold above background
80
60
40
20
0
SYBR Safe Stain
EtBr
TA97a TA98 TA100 TA102 TA1535 TA1537 TA1538
Ames test strain
SYBR Safe DNA Gel Stain
SYBR Safe DNA gel stain vs. ethidium bromide—Ames test results. Tests
were performed by Covance Laboratories Inc., Vienna, VA.
Summary of Results
A single oral administration of SYBR Safe DNA gel stain in
0.5X TBE at a limit dose of 5000mg/kg to three female rats
produced no mortalities or toxic signs. The procedure is designed
to determine that acute oral toxicity of the material under test.
A Limit Screen test was performed using three female Sprague
Dawley rats, which received an oral limit dose of 5000 mg/kg of
SYBR Safe DNA gel stain. The animals were observed for mortality,
weight change, and toxic signs for a two-week period. Since all
three rats survived for two weeks after the dose administration,
the LD50 for the test article was considered to be greater than the
Limit Dose and no additional testing was required.
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What’s the Worst That Could Happen?
Environmental Impact Results (USA)
Conclusion
SYBR Safe DNA gel stain is not classifi ed as corrosive, ignitable,
or reactive under the guidelines of the Environmental Protec-
tion Agency (EPA).
Table 3. SYBR Safe stain—Hazardous waste analysis
Analysis * Method Results
Corrosivity EPA 150.1 Not corrosive (pH = 8.25)
Corrosivity (by Corrositex) DOT-E 10904 Category 2 noncorrosive
Ignitability EPA 1010 Not ignitable (

Conclusion
SYBR Safe DNA gel stain in 0.5X TBE is indistinguishable from
0.5X TBE alone in terms of organic pollutant content—both
samples tested negative for the presence of an extensive
array of organic compounds.
Table 5. SYBR Safe stain—Detailed organic compound analysis *
Organochlorine pesticides and PCBs (608M)
alpha-BHC beta-BHC gamma-BHC delta-BHC (Lindane) Heptachlor
Aldrin Heptachlor Epoxide Endosulfan I Dieldrin 4,4’-DDE
Important Licensing Information
These products may be covered by one or more Limited Use Label Licenses (see the Invitrogen website,
www.invitrogen.com, or the Molecular Probes website, probes.invitrogen.com). By use of these products
you accept the terms and conditions of all applicable Limited Use Label Licenses. All products are for
research use only. CAUTION: Not intended for human or animal diagnostic or therapeutic uses.
SYBR Safe DNA Gel Stain
Endrin Endosulfan II 4,4’-DDD Endrin Aldehyde Endosulfan Sulfate
4,4’-DDT Toxaphene Chlordane Aroclor 1016 Aroclor 1221
Aroclor 1232 Aroclor 1242 Aroclor 1248 Aroclor 1254 Aroclor 1260
Ethylbenzene Bromoform 1,1,2,2,-Tetrachloroethane 1,3-Dichlorobenzene
Volatile organic compounds (624)
Chloromethane Vinyl Chloride Bromomethane Chloroethane Trichlorofl uoromethane
1,1-Dichloroethene Methylene Chloride trans-1,2-Dichloroethene 1,1-Dichloroethane Chloroform
1,1,1-Trichloroethane (TCA) Carbon Tetrachloride Benzene 1,2-Dichloroethane (EDC) Trichloroethene (TCE)
1,2-Dichloropropane Bromodichloromethane 2-Chloroethyl Vinyl Ether trans-1,3-Dichloropropene Toluene
cis-1,3-Dichloropropene 1,1,2-Trichloroethane Tetrachloroethene (PCE) Dibromochloromethane Chlorobenzene
1,4-Dichlorobenzene 1,2-Dichlorobenzene Acrolein Acrylonitrile
Semi-volatile organic compounds (625)
N-Nitrosodimethylamine Bis(2-chloroethyl) Ether Phenol 2-Chlorophenol Bis (2-chloroisopropyl) Ether
Hexachloroethane N-Nitrosodi-n-propylamine Nitrobenzene Isophorone 2-Nitrophenol
2,4-Dimethylphenol 4-Nitrophenol 2,4-Dichlorophenol 1,2,4-Trichlorobenzene Naphthalene
Hexachlorobutadiene 4-Chloro-3-methylphenol Hexachlorocyclopentadiene 2,4,6-Trichlorophenol 2-Chloronaphthalene
Acenaphthylene Dimethyl Phthalate 2,6-Dinitrotoluene Acenaphthene 2,4-Dinitrophenol
Bis (2-chloroethoxy) methane 2,4-Dinitrotoluene Fluorene Diethyl Phthalate 2-Methyl-4,6-dinitrophenol
N-Nitrosodiphenylamine 4-Bromophenyl Phenyl Ether Hexachlorobenzene Pentachlorophenol Phenanthrene
Anthracene Di-n-butyl Phthalate Fluoranthene Benzidine Pyrene
Butyl Benzyl Phthalate 3,3’-Dichlorobenzidine Benz(a)anthracene Chrysene Bis(2-ethylhexyl) Phthalate
Di-n-octyl Phthalate Benzo(b)fl uoranthene Benzo(k)fl uoranthene Benzo(a)pyrene Indeno(1,2,3-cd)pyrene
Dibenz(a,h)anthracene Benzo(g,h,I)perylene 1,2-Diphenylhydrazine 2,3,7,8-Tetrachlorodibenzo-p-dioxin 4-Chlorophenyl Phenyl Ether
* Samples analyzed were 0.5X TBE and 0.5X TBE + 1X SYBR Safe stain; none of the compounds listed in the table were detected in either sample. Testing was performed by Columbia Analytical
Services, Inc., Kelso, WA. Methods used were as outlined in the Code of Federal Regulations (CFR) Title 40, Part 136.
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