how safe is your gel stain? · a. redsafe™ b. safeview™ classic c. midori green advance figure...

How Safe is Your Gel Stain?

If it gets into living cells, is it really safe?

GelRed™ & GelGreen™ Nucleic Acid Gel Stains

• Non-toxic&non-mutagenic• Non-hazardousforeasydisposal• Ultra-sensitive• Stableatroomtemperature• Useinprecastgelorasapost-stain• Compatiblewithstandardinstruments• Compatiblewithcloningandsequencing• GelRed™replacesEtBr• GelGreen™replacesSYBR®Safe,EZ-Vision®RedSafe™,MidoriGreen,GreenSafe,SafeView™Classic,andothers

Acridineorange,ohnooo!

ManyresearchersseeksaferalternativestoethidiumbromidefortheirDNAgels.Anumberofdyeshavebeenmarketedasbeingsafebecausetheyarenotmutagenicinlabtests.Butmostoftheseso-calledsafegelstainscontainwell-known,old-fashionedfluorescentdyes,manyofwhicharehighlymembranepermeable,sotheypenetrateandbindDNAinsideoflivecellswithinminutes.Someofthesedyes,likeacridineorange,areknowntobehighlytoxictocells,andalsocanpotentiateDNAdamagebyUVlightandothermutagens.SeeTable1,otherside,foracomparisonofthecellpermeability,toxicity,andsensitivityofGelRed™andGelGreen™withotherethidiumbromidealternatives.

GelRed™andGelGreen™weredesignedspecificallytobecellmembraneimpermeable,andthereforenon-toxicandnon-mutagenic.Theyareclassifiedasnon-hazardouswasteunderCaliforniaTitle22.Inaddition,theyofferhighersensitivityandlowbackgroundcomparedtocompetinggelstains.

Formoreinformationandreferences,downloadourwhitepaper,ComparisonofNucleicAcidGelStains:CellPermeability,Safety,andSensitivityandthecompleteSafetyReportofGelRed™andGelGreen™atwww.biotium.com.

D.GreenSafePremium E.SYBR®Safe F.Nancy-5205min.

A.RedSafe™ C.MidoriGreenAdvanceB.SafeView™Classic

Figure1.Acridineorange-andSYBR-basednucleicacidgelstainsrapidlypenetratelivecellstostainthenucleus,cytoplasm,andotherorganelles.HeLacellswereincubatedwiththeindicatedgelstainincellculturemediumatthemanufacturer’srecommendedconcentrationforgelstainingforfiveminutesat37°C.Scalebars:20um.

A.GelRed™ A.GelGreen™Figure2.GelRedandGelGreendonotstainthenucleusinlivingcellsafter30minutesincubationat37°C.HeLacellswereincubatedwith3XGelRedor3XGelGreennucleicacidgelstainsincellculturemedium.InsetsshowCF640RWGAstaining(magenta)ofthesamefieldofviewtovisualcelloutlines.Brightstainingwasobservedonlyforsporadicdeadcellsontopofthecellmonolayer.A.GelRed.B.GelGreen.Scalebars:20um.

Many so-called safe gel stains rapidly penetrate living cells

GelRed™ and GelGreen™ are designed to be excluded from living cells

TryafreesampleofGelRed™orGelGreen™Emailorder@biotium.comorcall800-304-5357

OutsidetheUS,visitwww.biotium.comtofindadistributor

www.biotium.comGeneral Inquiries: order@biotium.com

Technical Support: techsupport@biotium.comPhone: 800-304-5357

Product Manufacturer Cat. No. Dye identity* Cell permeable? Dye toxicity* Sensitivity**

GelRed™ Biotium 41003 GelRed No Non-toxic 0.625-12.5ng

GelGreen™ Biotium 41005 GelGreen No Non-toxic 0.625-1.25ng

EZ-Vision®In-GelDye AMRESCO N391 DAPI No Non-toxic 5ng

SafeView™FireRed AppliedBiologicalMaterials G926 Propidiumiodide No Toxictocellswithprolongedexposure 0.625-1.25ng

RedSafe™ iNtRONBiotechnology 21141 Acridineorange Yes Toxictocells,potentiatesothermutagens 2.5ng

SafeView™Classic AppliedBiologicalMaterials G108 Acridineorange Yes Toxictocells,potentiatesothermutagens 2.5ng

MidoriGreenAdvance NipponGenetics MG04 Acridineorange+DAPI Yes Toxictocells,potentiatesothermutagens(acridineorange) 2.5ng

GreenSafePremium NZYTech MB13201 Acridineorange+DAPI Yes Toxictocells,potentiatesothermutagens(acridineorange) 2.5ng

SYBR®Safe ThermoFisherScientific S33102 Thiazoleorangederivative Yes Toxictocellsatworkingconcentration 2.5ng

Nancy-520 Sigma-Aldrich 01494 SYBR®derivative Yes Nodata 1.25ng

*Fordetailsofchemicalanalysisanddyetoxicityreferences,downloadtheComparisonofNucleicAcidGelStains:CellPermeability,Safety,andSensitivityatwww.biotium.com.**Detectionof500bpdsDNAfragmentbygelelectrophoresis.

Table 1. Comparison of non-mutagenic ethidium bromide alternatives

A.GelRed™ B.GelGreen™200100502512.5 200100502512.5

F.EZ-Vision®200100502512.5 200100502512.5

G.GreenSafePremium200100502512.5H.MidoriGreenAdvance

200100502512.5I.Nancy-520

200100502512.5J.SYBR®Safe

C.SafeView™FireRed200100502512.5

E.SafeView™Classic200100502512.5

D.RedSafe™200100502512.5

500bp-500bp-

500bp-

500bp- 500bp-

500bp-500bp- 500bp- 500bp- 500bp-

1052.51.250.625ng

GelRed™ and GelGreen™ also have superior sensitivity

Figure3.Comparisonofnucleicacidgelstainsbygelelectrophoresis.Biotium’s1kbDNAladderwasloadedonagarosegelsintwo-folddilutionsrangingfrom200ngto12.5ngtotalladderperlane.A.GelRedprecastgel.Themassofthe500bpbandineachlaneislabeled.B.GelGreenprecastgel.C.SafeViewFireRedprecastgel.D.RedSafeprecastgel.E.SafeViewClassicpost-stainedgel.F.EZ-Visionpost-stainedgelG.GreenSafePremiumpost-stainedgel.H.MidoriGreenAdvancepost-stainedgel.I.Nancy-520post-stainedgel.J.SYBRSafepost-stainedgel.

GelRed,GelGreen,andtheirusesarecoveredbyUSandinternationalpatents.EZ-VisionisaregisteredtrademarkofAMRESCO;SafeViewisatrademarkofAppliedBiologicalMaterials;RedSafeisatrademarkofiNtRONBiotechnology.SYBRisaregisteredtrademarkofThermoFisherScientific.