Lab Knowledge Base

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Revision as of 00:43, 9 May 2008 by Joy (Talk | contribs)

Contents

News

Wiki

  • Welcome to the Lab Wiki for Mark Farmer's Lab. This is the place to find procedures, news, and answers to commonly asked questions.
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Cultures & Lab Announcements

Professional Enrichment

  • The Association of Women in Sciences (AWIS)


Pressing Things TO DO!

Next Lab Meeting (?)

Today (08Apr08)

Things that must be done today:

  • refill agarose tubes
  • put dried volumetric pipets that are currently under paper towels by CC's desk space into metal containers and autoclave
  • wash dishes, especially the blue capped test tubes
  • autoclave the cleaned blue-capped test tubes and put at the bottom of the live-culture media/bottle storage shelf only to be used for bacterial work (i.e. not protist transfers)

If not finished, the last person to leave should e-mail SEJ or CC what's left to be done (depending on who it's for).

  • Joy, have you finished putting the labels on all of the test tubes ST transferred before he left? (he did all the racks with contaminants too...so double check that all of the latest transfers have labels on them) done JI
  • Inoculate 2 large flasks of ATCC 802 w/ Kpn and Ear. Incubate them overnight in 5B.Done ST
  • Stephen - Check the babies in the white rack in 1B. Spin down the ones that are ready.Checked, did not spin down ST
  • update UGA flasks sheet as to what we have in flasks...there were a bunch of small flasks at the bottom of incubator 4 that were completely dried down and never transferred to big flasks, so SEJ tossed these and they shouldn't be on the list now unless there are big flasks for them
  • Start new small flasks for these organisms if we don't already have them: updated UGA flask xls.; put all new small flasks - 4A
    • ECA dead in t.t.
    • EAN done CS;...redo b/c contaminated with other green euglenids/ciliates & maybe others...check orig. tube b4 transfer (SEJ 08May08)
    • EGY done CS; ...seems ok although maybe a small clear flagellate (SEJ 08May08)
    • EHS done CS
    • EGR done CS
    • MUT contamination w/ rotifers in t.t.
    • ESN done CS
    • ESL done CS
    • UEA dead in t.t.
    • ELA no mov't, all dead in t.t.
    • DAA done CS
    • CSK done CS
    • PYR done CS, maybe contaminated?
    • CCM contaminated w/ rotifers and dead in t.t.
    • CUA done CS


Priority:

  • back-up rack transfers
  • clean more blue-capped test tubes that SEJ autoclaved this morning (by live side sink)
  • make another set of LB+kan plates (follow directions on wiki) and be sure to store finished plates in walk in refrigerator with the solidified agar side up so condensation doesn't contaminate and drip onto the agar. (you will need at least 3-4 consecutive hours to do this or someone to immediately finish up where you leave off.)
  • need more CCAP S/Wdone but needs to be autoclaved and pHed BP
  • Empty all tip/trash box(s)/containers on gel and molecular benches every Wed & Fri
  • Wipe down all molecular work benches with the FIX 70% Ethanol squirt bottle every Wed & Fri



Cultures:

  • When checking cultures, please put aside any cultures with fungus for BP fungal treatments. Let him know where they've been put.
  • Brandon will be doing fungal treatment experiments using Sharan's notes in Lab Binder
    • Need to make 2 small flasks (one labeled microscopy) of the following (indicates test tube locatio); grow up small flask to be put in a large flask:
      • LUV (4C-backup)test tube contaminated with rodifers; used older test tube to make small flask; need to watch for contamination
      • TCI (4g)done CS
      • LXF (1g)done CS
      • LXI (4c-backup)done BP
      • LXJ (9h)test tubes contaminated with ciliates
      • LXH (1f)done CS
      • TUB test tubes contaminated
      • TUO JI did small flask (currently in 4), but seems like two different sizes of Trachs, redo small flask from uncontaminated tube or single cell isolate to get both types of cells isolated and grown up in small flasks done CS
      • SNB done CS
      • THB done CS
      • LUT (13d) done CS
      • TUN (?) can't find
      • TUK done CS
      • LUF (8f) done CS
      • LXR (6c) done CS
      • TUX done CS
      • LXT done CS
      • LYI (2d) done CS
      • LUK (9a) done CS
      • LYE (7f) done CS
    • Need to make 1 small flask for DNA (indicated test tube location):
      • LYA (7C)Contaminated with cilliates + rodifers; made a small flask - need to watch for contamination
      • SXD (4i) Dead in notebook 30Jan08
      • LXL (1d) done CS
      • SXE (5f)done BP
      • TSP (10b)done BP
      • TUY (3b)done BP
      • TXB (4d) done CS
      • TXG (1c) done CS...please do again b/c no trach cells on 26Feb08, only ciliates in small flask test tubes contain no living cells or ciliates; may have 1 cell in test tube 4 Oct 07 - did not make small flask
      • LXW (6f) done CS
      • LXX (1i) done CS
      • LYB (?) Can't find
      • LUW (9b) done CS
      • LUO (4c-backup) can't find CS;This was thrown out 30Jan08, but we still have a good big flask for it- JI
      • LYS (13j done CS
      • LYT (13i) done CS
      • LUA (13h) done CS
      • LUC (13g) done CS
      • LUD (13f) done CS
      • LUG (13e) done CS
      • LYP (?) Can't find
      • LYD (7e) done CS
      • LYG (7h) done CS
      • LXN (?) Can't Find
      • SUM (?) Can't Find
      • TVL test tubes contaminated
      • LYU test tubes contaminated
      • LXW done CS
      • LUW done CS
      • LXZ done CS
      • LXP done CS
      • LUU done CS
      • SUL done CS


Lab Assistants, Please Update Sarah/Christina On:

  • Updates for Sarah
    • list of all cultures I need in small flasks that have been started (any moved to big flasks yet?)
    • list of all cultures I need in small flasks that are contaminated (and preferably what they are contaminated with)
  • Updates for Christina
    • A small flask of EGY is located on 4C and maybe contaminated with one of your organisms 14Apr08 CS



Notes for Sarah

  • CS, please spin down TET for me on 18Apr08, if not done already, and put it in the walk-in fridge, but let me know when it is ready (SEJ - posted 17Apr08)done CS in -80 freezer; cell box 4-H1
  • postponed until we have tried the RNA-cDNA process...SEJ's - Please do DNA extractions on the following organisms (be sure to check if they original spin down was done on Log phase cells before you combine/extract them):
    • Cell Box 2: SBR, STB, TZR, THA, TPL, TUM, TVISBR done BP
    • Cell Box 3: LXS, SNB, LUO, SXA, SSA, MUC=MUR, SSC
    • Cell Box 4: ELU, SMA, SUZ, SEU, TUA
    • Cell Box 6: TBR, LUE, TVS, TRG, TUJ, SMC
    • Cell Box 7: SUN, CNT, TUX, TUK, SUY, SBA,
    • Cell Box 8: SXB, SUI
    • Cell Box 9: TOG, TUV, TEP, TSP, SCP
    • Cell Box 10: TLF=TLV (some tubes are mislabeled as TLF when they should be TLV=Trachelomonas lefevrei)

Are you a Lab Assistant with free time on your hands?

  • Make any media that needs to be made (generally AF-6 Medium, ESNW or Hay Infusion)
  • Autoclave media using the liquid cycle (be sure you label with autoclave tape)
  • Using sterile technique, add any additional ingredients to complete media post-autoclaving (eg. vitamins)
  • Wash test tubes, media containers and their respective caps (keep LIVE and FIX glassware separated)
  • Autoclave the test tubes, media containers, 9" pasteur pipets, pipet tip box(es), volumetric pipets, etc. in appropriate containers with containers underneath if available
  • Using sterile techniqe, add media to the autoclaved tubes or sterile culture flasks
  • Check and make notes of cultures without opening the tubes or culture flasks using the appropriate microscopes. Some cultures need to be checked daily, weekly, bi-weekly or monthly depending on their growth patterns.
  • Make transfers of healthy cultures (all culture information can be found on the Cumulative Culture Inventory.xls sheet on the Farmer Lab PC desktop)
    • tubes
    • big/small flasks
  • Spin down log phase cells in big culture flasks and rejuvenate with fresh media (aseptically)
  • Extract DNA from concentrated cells and update the DNA.xls sheet
  • Qualify/Quantify DNA on Nanodrop
  • Maintain molecular buffers, reagents
  • Maintain at least one carboy of DI water
  • Keep everything clean and organized
  • Ask if anyone in the lab needs anything done



Lab Binder & Notebook

Phylogenetics and Systematics of Euglenozoa

Documents



See Also

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