Frequently Asked Questions

Questions

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►Can I re-use the plastic transfer pipettes that come with the KCK kit?
Rinse or soak in soapy water.  Pull soapy water in and out to clean inside.  Rinse/soak in plain tap water.  Again, pull water in and out to rinse inside.  Be sure to rinse the whole inside of the bulb.  Allow to dry or drain.  Finally rinse with 70% alcohol.  Pull alcohol in and out of pipet include the bulb.  Drain with tip side down on paper toweling.  They transfer pipettes are ready to use again.  They are NOT sterile but free of chemical contaminants.  Note that these sell for about $12-20 for 500 which is about 4 cents each.  If you can't find them locally, email me.  I think the weight of an envelope with a few of these inside should cost no more that 2 US stamps. 
►Is distilled water that we purchase from the store "sterile" and ready to use for rinsing explants?
No.  While it might have been sterile when processed, it is no longer sterile once you pour it from the bottle into another container.  Distilled water must be processed in a microwave or pressure cooker before it can be used as a "sterile water" rinse.
►How do I rinse my explants after they have soaked in the dilute bleach solution?
Prepare sterile water as described in the KCK handout or KCK Manual.  After containers are cool, move one or more to the cleanbox.  Be sure to spray the inside of the cleanbox with 70% alcohol or dilute bleach (see handout).  Place container with explants and bleach solution in the hood.  Spray that with alcohol too.  Using a disinfected forceps, pick up explants and place in the sterile water.  Allow to soak for a few minutes.  Some protocols recommend doing this several times. If so, move the explants (with a disinfected forceps) to a second container of sterile water.  Explants can usually soak in the water for several minutes" rinse before being trimmed and then cultured on sterile medium.
►What procedures are done outside of the cleanbox, and which ones are done inside the cleanbox?
Preparing the explant for culture is done outside the cleanbox until you put the explant into the bleach or alcohol solution.  Then it needs to be moved to the cleanbox where it will be protected from dust and microorganisms.
►How do I store the media once it has been processed in the microwave or pressure cooker?
Once the culture vessels with sterile medium are cooled, they should be stored  in plastic boxes with covers or on trays inside of ziplock bags.  Alternatively each could be wrapped with plastic wrap.  All of these methods will keep the dust out of the jars and slow dessication (drying) of the media.  Store these at ROOM TEMPERATURE.  DO NOT REFRIGERATE.
►What other glass jars can I use besides baby food jars?  What do I use for covers if I plan to microwave these jars?
Glass jars used for jam, spaghetti sauce, pickles, etc. are the same size at the Unicorn #2 cover that we sell and can be used.  You can use those when you microwave the media and then switch to original metal cover in the clean box.  The original metal cover can be disinfected with alcohol.  The unicorn cover can be used for the life of the culture.  Wrap with plastic wrap to keep it secure.

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