GoodsteriletechniqueisthefirstandmostimportantstepininsuringconsistentresultswhenemployingrecombinantDNAandproteinexpressiontechniques.Steriletechniquereferstoproceduresbywhichculturesmaybemanipulatedwithoutinfectingtheworkerorcontaminatingtheculturesorthelaboratoryenvironment.Becausecontaminatingbacteriaareubiquitousandarefoundonfingertips,benchtops,etc.,itisimportanttominimizecontactwiththesecontaminatingsurfaces.Whenstudentsareworkingwiththeinoculationloopsandagarplates,youshouldstressthattheroundcircleattheendoftheloop,thetipofthePipetter,andthesurfaceoftheagarplateshouldnotbetouchedorplacedontocontaminatingsurfaces.TheflamingoflipsoftubesandflasksmustALWAYSbedonewhenevercultureliquidistobepouredfromacontainer(e.g.,pouringplates).Flamingshouldberoutinelydonewhencapsareremovedfromtubesduringtransferofcultures.Thepurposeofflamingisnottosterilize,buttowarmthetubeandcreatewarmairconvectioncurrentsupandawayfromtheopening.This"umbrella"ofwarm,risingairwillhelptopreventtheentranceofdustparticlesuponwhichcontaminatingbacteriareside.Petridishlidspreventdustfromfallingdirectlyontoplatesbutallowdiffusionofairaroundtheedges.Therearenodirectaircurrentsintotheplate,andtoenter,dustparticleswouldhavetoriseverticallymorethanacentimeter.Thisdoesnotoftenoccurbecauseofthedensityoftheparticles.Wheneverthelidisremoved,itshouldbeheldovertheplateasashield.Donotplacethelidonthebenchtop.Donotleaveplatesuncovered.Donotwalkaroundtheroomwithanopenplate.Whenworkingwithculturesintesttubes,workasrapidlyasisconsistentwithcarefultechnique.Keepthetubesopenaminimumamountoftime.Whilethetubesareopen,holdthemata45degreeanglesothatdustcannotfallintotheopentube.Holdthetubesawayfromyourfacewhiletransferring.Testtubesarehandledinthefollowingmanner:

• Thetesttubeisheldinthelefthand(foraright-handedperson).
• Theinstrument(loop,pipet,orneedle)isheldintherighthand.
• Thetesttubecapisgraspedbythelittlefingeroftherighthand,andremoved.
• Whilecontinuingtoholdthecapwiththelittlefinger,thetubeislightlyflamedandtheinstrumentismanipulatedappropriately,andwithdrawn.
• Thecapisreplacedonthetesttubeandthetesttubeisputbackintotherack.

Labelallcultureswiththenameornumberoftheorganism,andyourname.Alwayscleanallworkareas(yourbench,balancearea,sinkarea,gelarea,etc.)thoroughlybeforeleavingthelaboratory!Thelaststepbeforeleavingthelabistowashyourhandsthoroughly.Theseareguidelines.Youmayfindasetoftechniquesthatbestsuiteyourworkingstyle.Thisisfineaslongasyouadheretothebasicconceptsofgoodsteriletechnique.

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