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Pouring platesEdit

  • Pour plates the week before the RLS is to be started. Pour 28ml / plate.
  • Use Falcon brand REF 351029 10 cm plates, as not all 10 cm plates fit the dissection scopes properly.
  • Leave the plates out to dry for at least the 2 days before the RLS is started. Wet plates make dissection difficult.

Getting cells readyEdit

  • Pull cells from -80C to YPD and let grow to colonies 2-3 days or as needed. Typically pull Thurs for the Monday RLS and Friday for the the Tuesday RLS.
  • Typcially, one prepatch is done to the same media that will be used for disection conditions. Pick cells that are representative from the YPD plate for the prepatch. If the strain is slow growing, don't pick a colony that may contain a suppressor mutation, e.g., faster growing which will usually be a large colony among many smaller colonies.
  • Patching (Note: do not perform without being specifically trained for patching )
    • Patch a second time to plates which will be used for the RLS assay. Typically 3 patches per plate (each patch will be used for 20 cells RLS) are done. Match the strain # from the lifespan printout sheet to each ID # for the RLS. Make sure to patch the correct strain to the correct number and also to the correct media.  Wrap plates in parafilm and place into the back of the 30C lifespan incubator and be sure to also write the RLS # on each plate. The dissection team will know what to do from here. If the cells are setup differently from normal, make sure to be there to instruct during the setup process.
    • Always make sure to use sterile toothpicks (these should have autoclave tape somewhere on them, usually the lid, and black lines in the autoclave tape indicating sterility). Be sure you are not using used toothpicks.
    • If there are any significant questions or confusion, call or email the person who submitted the lifespan and get clarification (see the contact sheet for phone numbers).

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