Lab Knowledge Base

Revision as of 15:47, 26 September 2007 by Christina (Talk | contribs)

News

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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Some of my lovely babies are growing!!! (Hooray!!) The Rhynchobodo sp. (RBS) are growing in a small flask in the 25C incubator. Please feel free to take a look at these new children of the lab. Also, please take a moment to mourn the loss of the other 2 ATCC cultures that came in (Diplonema ambulator and Trypanoplasma borreli). They did not survive past birth.
ELU prefers to be at room temperature (RT) in the saltwater media (MSU), but K media works too (back up tubes & flasks in 20C)
ETM (15C) and ELG (20C) also seem to prefer saltwater media (MSU), but K media works too
WE HAVE A -80 FREEZER NOW!!! (we need to get some crates or something to put in there for now, but then you can start storing our DNA and cells in there!)
be sure to take scrupulous notes when doing the DNA extractions (cell tube info, extraction info, nanodrop curve info, etc.)

The Association of Women in Sciences (AWIS)
- About the Organization: AWIS brings together undergraduate and graduate women in the sciences to discuss science, careers and lifestyles in science as well as providing organized opportunities for science outreach in the local community. There are no dues and both men and women are encouraged to join! Contact Sana Hashmi (AWIS Undergraduate Presidentt) to be added to the AWIS listserve.
- Outreach:
  - Dates:
    - September 27th, 3:30-4:30pm
    - October 18th, 3:30-4:30pm
    - November 15th, 3:30-4:30pm
  - Where: Oglethrope Elementary School (carpools are available)
  - What: helping elementary students conduct small science experiments
- First Seminar of the Semester:
  - When: time TBA, October 2nd
  - Where: TBA
  - Who: Dr. Maxia Dong, Center for Disease Control (CDC) and the National Institute of Health (NIH)
  - What: TBA

Priority:

Please cut and attach another piece of styrofoam to the broken ended one in the -80C freezer (make sure it still sits level)
Please glue gun the leak in the pipette cleaner next to the Nanodrop area.
move stuff from molecular/PCR area and clean off top shelves...wipe of lower bench with ethanol when done and then replace all the stuff that was moved off temporarily (check with Tina and Sarah first to be sure they don't need to work on the bench while you are doing it)

Cultures:

Spin down the 3 large flasks of PCT (20C). Make sure to scrape the bottom of the flask with a sterile rubber policeman. Dump the flasks afterwards.
spin down cells in big flasks (check Caitlin's notes from thursday)
Brandon will be doing fungal treatment experiments using Sharan's notes in Lab Binder

Transfers:

start growing any other MSU cultures up in a small flasks (that haven't been done already) in addition to the back up tube (recently added to the culture inventory...some might already be in their appropriate rack number if they are just replacing a culture we had previously)...will need lots of AF6
DMT (25C) - Scrape the bottom of the 2 large flasks with a sterile rubber policeman. Dump the majority of the media. Refresh the media with the 2 large flasks of ATCC 802 w/ Kpn and Ear in the 33.5C incubator.
RBS (25C) - Scrape the bottom of the 3 small flasks with a sterile rubber policeman. Distribute the majority of the media to the 2 large flasks of ATCC 802 w/ Kpn and Ear in the 33.5C incubator. Refresh the media in the small flasks with the media in the 3 small flasks of ATCC 802 w/ Kpn and Ear in the 33.5C incubator.
Kpn and Ear (33.5C) - Inoculate 2 large flasks of ATCC 802 w/ Kpn and Ear (in 4C walk-in). Grow on the middle shelf of the 33.5C incubator.
PCT (20C) - Agitate the 6 small flasks. Dump half of the media. Replace with fresh ESNW and 2-3 new grains of sterile rice.

Media:

Lab Assistants, Please Update Sarah/Tina On:

check back up small flasks & "no cells" small flasks in the culture room 20C incubator (transfers made or back ups checked & ok?)

Other:

update chemical inventory with the chemicals whose MSDS sheets are next to the Farmer Lab PC and then put the MSDS sheets away alphabetically in the MSDS binder
drill two extra holes in the black drying rack for the culture room and mount it on the wall above the current dishwashing area (move all glassware and dishwashing items first so they don't get broken or in the way)...at least 2-3 people will be needed to lift and mount the drying rack
update culture inventory with which racks recently added MSU cultures are actually in (edit if some are repeats)
wash dishes (make sure dry before putting away!)
set up new answering machine to lab phone (with personalized message!)
when grown up, pour all of T411 small flask & large flask into centrifuge tubes and spin down (add 15mL of AF6 back to small flask)...green algal cells only, so be sure not to contaminate any of our other cultures with them!!!

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
- tubes
- big/small flasks
Spin down log phase cells in big culture flasks and rejuvenate with fresh media (aseptically)
Extract DNA from concentrated cells
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