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PetakaG3 TMBasicsMaximum volume of media permittedin Petaka isMaximumvolume20 mlMedia injection and withdrawalshould always be done holdingPetaka in a VERTICAL POSITION(more will alter Petaka’s performance!)PetakaG3 TMReady to hostPetakas (RTHP)PREPARING “READY TOHOST” PETAKASHold the Petaka in a vertical position!1. Inject 0.5 ml of Antibiotic solution2. Pump 16 ml of MediumTotal16.5mlKeep 3.5 mlfree to addcells and serumlater3. Keep these Petakas refrigerated at4ºC in a vertical position in standsor in a plastic bag (zip bag)Dehydration and pH should becontrolled if RTHP are kept Inrefrigerator for months!SEEDING CELLS IN “READYTO HOST” PETAKASHolding Petaka a in vertical position1. Disinfect port with 90% Ethanol &a flame2. Warm the Petakas in an incubator,at 37ºC for 20 -30 min in aVERTICAL position3. Inject 1.5ml of serum (*)4. Inject up to 2ml of cell suspension5. Whilst holding the Petaka in avertical position, squeeze it untilthe level of media reaches the airexit & then release until the air inputstops. Then the inside/outsidepressure will be balanced6. Shake gently for 10-15 seconds7. Place in a stand & allow to incubatein a HORIZONTAL positionCulturing cells in mediasupplemented mediaPetakaG3 TM mlSEEDING THE CELLSHolding Petaka in a vertical position!1. Inject 1 ml of Serum for 5%concentration2. Inject 0.5 ml of Antibiotic solution3. Inject X ml of Cell suspension4. Inject (18.5 –X) ml of Medium20Total5. Whilst holding the Petaka in avertical position, squeeze it untilthe level of media reaches the airexit & then release until the air inputstops. Then the inside/outsidepressure will be balanced.6. Incubate Petaka according tothe Cell type (see INCUBATIONPOSITIONS)