LESSON 8: INOCULATION AND STREAKING TECHNIQUESSON ##: Lesson TitleINTRODUCTIONAs a microbiology student, you will be required totransfer living microbes from one place to anotheraseptically (i.e., without contamination of theculture, the sterile medium, or the surroundings).While you won’t be expected to master all transfermethods right now, you will be expected to performmost of them over the course of the semester. Referback to this section as needed.Aseptic transfersare not difficult; however, a littlepreparation will help assure a safe and successfulprocedure. Before you begin, you will need to knowwhere the sample is coming from, its destination,and the type of transfer instrument to be used.These exercises provide step-by-step descriptions ofdifferent transfer methods. In an effort to avoid toomuch repetition, skills that are basic to mosttransfers are described in detail once under “TheBasics” and mentioned only briefly as they apply totransfers in the discussion of “Specific TransferMethods.”ASEPTIC TECHNIQUEThe use of aseptic technique insures that nocontaminating organisms are introduced into culturematerials when the latter are inoculated or handledin some manner. It also insures that organisms thatare being handled do not contaminate the handler orothers who may be present. And its use means thatno contamination remains after you have workedwith cultures.To gain some practice in aseptic transfer of bacterialcultures, three simple transfers will be performed inthis Lesson: (a) broth culture to broth tube, (b) agarslant culture to an agar slant and (c) agar plate to anagar slant.NOTE!To prevent contamination of the sample,inoculating instruments must be sterilized prior touse. Inoculatingloops and needles are sterilized immediatelybefore use in an incinerator or Bunsen burnerflame. The mouths oftubes or flasks containing cultures or media arealso incinerated at the time of transfer by passingtheir openings through a flame. Instruments thatare not conveniently or safely incinerated, such asPasteur pipettes, cotton applicators, glasspipettes, and digital pipettor tips are sterilizedinside wrappers or containers by autoclaving priorto use.MATERIALS AND EQUIPMENTTable 1.Types of Transfer and Materials &EquipmentTYPES OF TRANSFERMATERIALS &EQUIPMENTBroth culture to Brothtube●Broth culture●Tubes of sterilenutrient broth●Inoculating loop●Bunsen burner orIncineratorAgar slant culture toAgar slant●Agar slant culture●Sterile nutrientagar slant●Inoculating loop●Bunsen burner orIncineratorAgar plate to Agarslant●Nutrient agar platewith bacterialcolonies●Sterile nutrientagar slant●Inoculating loop●Bunsen burner orIncineratorLABORATORY PROCEDURETRANSFERRING FROM BROTH CULTURE TOANOTHER BROTHPREPARED BY: ESTHER VICTORIA H. TOLENTINO & RICCI ANNE V. LAUREZO | BSMT 3-31OUTLINEIAseptic Technique and Inoculation MethodsA.Materials and EquipmentIILaboratory ProcedureA.Transferring from Broth Culture to Another BrothB.Transferring from a Slant Agar Culture to Sterile NutrientAgar SlantC.