Designation:J99DepositedName:Helicobacterpylori(Marshalletal.)Goodwinetal.Genotype:cagA+vacA+ProductDescription:Genomesequencingstrain.
MediumATCCMedium18:TrypticaseSoyAgar/BrothATCCMedium260:Trypticasesoyagar/brothwithdefibrinatedsheepblood
GrowthConditionsTemperature:37CAtmosphere:Microaerophilic,35%O2,10%CO2
PropagationProcedure1.
Thisorganismisshippedfrozenindryice.Justpriortouse,thawvialinwateratapproximately
37C.Whenthawed,adropofthesuspensionmaybeusedtodoanimmediatewetmounttoobservetheuniquemorphologyofthisorganismandverifyitsviabilitybycheckingformotility.
2.
Asepticallytransferthethawedsuspensionintoafresh#18broth(3to5mL).Mixwell.Thissuspensioncannowbeusedtoinoculateagarslant(s),plate(s),orthepreferredbiphasicculture.Two#260platesshouldbeinoculated,oneformicroaerophilicgrowthandthesecondforaerobicgrowth.Nogrowthshouldoccurontheplateincubatedaerobically.
3.
Toobtainabiphasicculture,add0.6mLofthesuspensiontoa#260slant.Theresultingpoolatthebottomoftheslantiswherethebest,mostrapidgrowthwilloccur.
4.
Incubateat37Cundermicroaerophilicconditionsusingananaerobejarwithanactivecatalystandamicroaerophilicgasgeneratorpack,orotheracceptablemethod,toobtainmicroaerophilicconditions.Incubatetubeswithcaploose.
5.
Within3days,goodgrowthshouldbeobtainedinthebrothpoolatthebottomoftheslant.Additionalincubationmayberequiredforcoloniestoappearonagarplate.Furthersubculturescanbemadeusingthebrothpoolastheinoculumsource.Subculturestobiphasiccultureswillrequireonly24to48hoursofincubationforgoodgrowth.
Thisisaslowgrowingorganismthatrequiresmoistconditionsforbestgrowth.Growthatthebroth/agarinterfaceofthebiphasicslantshouldoccurwithin3days,butonlylightturbiditywillbeseen.Toobservegrowth,examineawetmountofthebrothunderphasemicroscopy.Thisorganismisamediumsize,regulartoslightlycurvedtoweakspiral,motile,Gramnegativerod.
Growthonagartakeslongerthanthebiphasicculture.Coloniesaresmallsmooth,transparent,butyrous,circularandlowconvexwithanentireedge.Itisessentialtousefresh,moistplates.ThecellsdonotGramstainwellusingtraditionalprocedures.Forbestresults,useabasicfuchsincounterstaininplaceofthesafranin.
Oncegoodgrowthinobtained,transferorfreezetheculture.Addinganequalamountof20%sterileglyceroltopooledbrothfromseveralbiphasicslants,followedbyfreezinginliquidnitrogenorultralowtemperaturefreezerisrecommended.
PurifiedgenomicDNAofthisstrainisavailableasATCC700824D5.AdditionalinformationonthiscultureisavailableontheATCCwebsiteatwww.atcc.org.
Referencesandotherinformationrelatingtothisproductareavailableonlineatwww.atcc.org.
Appropriatesafetyproceduresshouldalwaysbeusedwiththismaterial.LaboratorysafetyisdiscussedinthecurrentpublicationoftheBiosafetyinMicrobiologicalandBiomedicalLaboratoriesfromtheU.S.DepartmentofHealthandHumanServicesCentersforDiseaseControlandPreventionandNationalInstitutes
Description
Propagation
Notes
References
BiosafetyLevel:2
Page1of2
ProductSheet
Helicobacterpylori(ATCC
700824)
PleasereadthisFIRST
StorageTemp.Frozen:80CorcolderFreezeDried:2Cto8CLiveCulture:SeePropagationSection
BiosafetyLevel2
IntendedUse
Thisproductisintendedforresearchuseonly.Itisnotintendedforanyanimalorhumantherapeuticordiagnosticuse.
CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,itshouldbecitedinthatmanuscriptinthefollowingmanner:Helicobacterpylori(ATCC700824)
AmericanTypeCultureCollectionPOBox1549Manassas,VA20108USAwww.atcc.org
800.638.6597or703.365.2700Fax:703.365.2750Email:[email protected]
forHealth.
ATCCWarranty
TheviabilityofATCCproductsiswarrantedfor30daysfromthedateofshipment,andisvalidonlyiftheproductisstoredandculturedaccordingtotheinformationincludedonthisproductinformationsheet.ATCCliststhemediaformulationthathasbeenfoundtobeeffectiveforthisstrain.Whileother,unspecifiedmediamayalsoproducesatisfactoryresults,achangeinmediaortheabsenceofanadditivefromtheATCCrecommendedmediamayaffectrecovery,growthand/orfunctionofthisstrain.Ifanalternativemediumformulationisused,theATCCwarrantyforviabilityisnolongervalid.
Disclaimers
Thisproductisintendedforlaboratoryresearchpurposesonly.Itisnotintendedforuseinhumans.WhileATCCusesreasonableeffortstoincludeaccurateanduptodateinformationonthisproduct
sheet,ATCCmakesnowarrantiesorrepresentationsastoitsaccuracy.Citationsfromscientificliteratureandpatentsareprovidedforinformationalpurposesonly.ATCCdoesnotwarrantthatsuchinformationhasbeenconfirmedtobeaccurate.
Thisproductissentwiththeconditionthatyouareresponsibleforitssafestorage,handling,anduse.ATCCisnotliableforanydamagesorinjuriesarisingfromreceiptand/oruseofthisproduct.Whilereasonableeffortismadetoinsureauthenticityandreliabilityofstrainsondeposit,ATCCisnotliablefordamagesarisingfromthemisidentificationormisrepresentationofcultures.
PleaseseetheenclosedMaterialTransferAgreement(MTA)forfurtherdetailsregardingtheuseofthisproduct.TheMTAisalsoavailableonourWebsiteatwww.atcc.org
AdditionalinformationonthiscultureisavailableontheATCCwebsiteatwww.atcc.org.ATCC2014.Allrightsreserved.ATCCisaregisteredtrademarkoftheAmericanTypeCultureCollection.[02/04]
Page2of2
ProductSheet
Helicobacterpylori(ATCC
700824)
PleasereadthisFIRST
StorageTemp.Frozen:80CorcolderFreezeDried:2Cto8CLiveCulture:SeePropagationSection
BiosafetyLevel2
IntendedUse
Thisproductisintendedforresearchuseonly.Itisnotintendedforanyanimalorhumantherapeuticordiagnosticuse.
CitationofStrain
Ifuseofthiscultureresultsinascientificpublication,itshouldbecitedinthatmanuscriptinthefollowingmanner:Helicobacterpylori(ATCC700824)
AmericanTypeCultureCollectionPOBox1549Manassas,VA20108USAwww.atcc.org
800.638.6597or703.365.2700Fax:703.365.2750Email:[email protected]
