Lab Knowledge Base

From Protists

(Difference between revisions)
(→Today (23Feb08))
(→Today (23Feb08))
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*Take the 2 plates out of 37C and put them in 4C walk-in next to the Kpn plates.  
*Take the 2 plates out of 37C and put them in 4C walk-in next to the Kpn plates.  
*Spin down and refresh the following large flasks: (Use sterile scrapers and do a quick spin to remove soil, wheat, and cerophyll)
*Spin down and refresh the following large flasks: (Use sterile scrapers and do a quick spin to remove soil, wheat, and cerophyll)
-
**RBS (6A)
+
**RBS (6A) '''done CS&BD'''
**NBD (6A)
**NBD (6A)
**DMT (6A) Make sure not to allow the media to be open in presence of DMT. Use centrifuge tubes for the media transfer to the flasks.
**DMT (6A) Make sure not to allow the media to be open in presence of DMT. Use centrifuge tubes for the media transfer to the flasks.
*Check to see if we need HI, ATCC 802, and CCAP S/W. If so, please start making more or put it on the wiki for others to do.
*Check to see if we need HI, ATCC 802, and CCAP S/W. If so, please start making more or put it on the wiki for others to do.
 +
**'''we need [[Hay Infusion]] (1L) and [[CCAP S/W]] (3L)
*Make Art. SW with 33.6g of Ultramarine salts in 1L of dH2O. Use stir bar and heat to mix. Let settle in appropriate carboy for 2 days before filtering and autoclaving.  
*Make Art. SW with 33.6g of Ultramarine salts in 1L of dH2O. Use stir bar and heat to mix. Let settle in appropriate carboy for 2 days before filtering and autoclaving.  
*check flasks and spin down if ready... (don't forget the flasks in the #2 and #3 incubators)...switch over any of Sarah's in closed flasks into vented flasks or start new small ones from scratch if original big flasks dead '''DO in progress'''
*check flasks and spin down if ready... (don't forget the flasks in the #2 and #3 incubators)...switch over any of Sarah's in closed flasks into vented flasks or start new small ones from scratch if original big flasks dead '''DO in progress'''

Revision as of 18:09, 23 February 2009

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.
  • If you are new to this wiki, you will need an account in order to edit pages. Talk to Sarah about getting an account set up if you would like to contribute.
  • You can upload pictures, word, excel, and pdf documents.
  • If you are new to wiki editing, check out the wiki help for how to edit articles in a wiki, then use the existing pages as templates and examples of how to create links, upload files, and display images.


Cultures & Lab Announcements

Professional Enrichment

  • The Association of Women in Sciences (AWIS)


Pressing Things TO DO!

Next Lab Meeting (20Feb09 @ 2pm!!!)

  1. go over spin down procedures for RP1, DIP & RBS
  2. Cell Boxes updated?
  3. Introductions for new labbies (birthdays on dry erase board)

Today (23Feb08)

Things that must be done today or tomorrow:

  • Take the 2 plates out of 37C and put them in 4C walk-in next to the Kpn plates.
  • Spin down and refresh the following large flasks: (Use sterile scrapers and do a quick spin to remove soil, wheat, and cerophyll)
    • RBS (6A) done CS&BD
    • NBD (6A)
    • DMT (6A) Make sure not to allow the media to be open in presence of DMT. Use centrifuge tubes for the media transfer to the flasks.
  • Check to see if we need HI, ATCC 802, and CCAP S/W. If so, please start making more or put it on the wiki for others to do.
  • Make Art. SW with 33.6g of Ultramarine salts in 1L of dH2O. Use stir bar and heat to mix. Let settle in appropriate carboy for 2 days before filtering and autoclaving.
  • check flasks and spin down if ready... (don't forget the flasks in the #2 and #3 incubators)...switch over any of Sarah's in closed flasks into vented flasks or start new small ones from scratch if original big flasks dead DO in progress


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).


Priority:

  • DNA extraction of SNB (see Sarah and CC to see if more need to be done too)
  • Joy, please start checking the test tube racks and start making transfers ASAP 6Feb09: Finished looking at racks, will start to transfer...(JI)
  • 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
  • keep up with washing, drying and autoclaving the glass volumetric pipets so we don't run out (5, 10, 25mL) done CS - just need to be autoclaved 20Feb09
  • put cotton in the tops of pasteur pipets and fill the containers (by the dishwashing station) and autoclave them Zg & JI 6Feb09
  • Empty all tip/trash box(s)/containers on gel and molecular benches every Wed & Fri done CS 20Feb09
  • Wipe down all molecular work benches with the FIX 70% Ethanol squirt bottle every Wed & Fri done CS 20Feb09
  • To do for JI: 1. Update cell inventory (done), 2. Make small flask of ELU (done Feb09 JI), 3. Make small flasks of old contaminated flasks (from test tubes) done


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 location); 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



Notes for Sarah

  • check blue rack on shelf D of incubator 3 with ELG/ELU/etc. in it...why haven't these been transferred since 2007? JI checked and none survived except ELU. I autoclaved/threw out dead tubes and made small flask of ELU. 6Feb09
  • 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)
  • make new GRC big flask done CS
  • 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...Made new small flask (4B) ST 9Jun08
    • EGY Made new small flask (4B) ST 9Jun08
    • 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

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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