Maintenance Tasks

General Upkeep (everyone)

All members of the lab are responsible for maintaining the communal lab environment. Everyone is responsible for cleaning spaces and equipment after use and storing reagents in their proper locations. This includes:

Wipe benches. Use 70% ethanol and a paper towel to wipe and clean personal spaces on a regular basis and common spaces after use.
Clean analytical balance area. Use 70% ethanol on a paper towel to clean chemical powders. Clean the inside of the balance and the area outside of it. Do not just brush powders onto the floor! Brush powders into a trash can.
Clean microscope lenses. Spray Sparkle (purple glass cleaner) on a Kimwipe and gently wipe the lenses of microscopes after use or as needed.
Clean sink area: ?
Biohazard bag decontamination. There are several red containers around the lab with autoclave bags in them. When these are ~75% full, take out the autoclave bag and tie it closed. Use lab tape to ensure it stays closed. Label the bag with our lab name and then place the bag in the MERI bin in Rm 2934.
Styrofoam: place styrofoam in dedicated hallway container. ?

General Reagents (assigned)

The items below occur as needed. Lab members should warn the assigned person several days in advance and write on the white board in Room 2951 when a task needs to be completed:

50x TAE and refilling 1x TAE carboy. Refill the 1x TAE carboy with milli-Q water from the carboy and 50x TAE. Make 50x TAE. See General Recipes for instructions.
Ordering and aliquoting PCR reagents (ladder, dye, dNTP stocks) Make 6x orange loading dye and aliquot into 1.5 mL tubes. See the loading dye recipe in General Recipes.See Ladder Creation for instructions to create ladders from plasmids or directly order and aliquot ladder. Combine individual commercial 100 mM dNTP stocks (dATP, dTTP, dCTP, dGTP) to make 10 mM working stock aliquots for lab member personal use. Add 10 μL of each dNTP to 60 μL of dH₂O to make 100 μL aliquots of 10 mM dNTP working stocks.
Ordering and aliquoting culture reagents Maintaing stocks of media (RPMI), antibiotics, and serum (FBS) for general parasite and mammalian cell culture.
Competent cells. See the Competent Bacteria protocol for instructions.
SOC. Make 100 mL of SOC media for lab use. Aliquot into 10 mL units and store at 4°C. See General Recipes for instructions.
K medium Make and filter 1 L stocks without cholesterol. Cholesterol should be added to the aliquot prior to use and used within 3 weeks of preparation.
Worm 2x lysis buffer. This needs to be made rarely, in large batches and frozen. See C. elegans Recipes.
M9 stock replenishment. Make 4X stock solutions of M9. Follow the recipe in C. elegans Recipes. Be sure to autoclave and then add 1 M MgSO₄ once cool.

General Tasks (assigned)

Take care of general lab waste. There are "Waste" and "Liquid Waste" plastic containers on benches. "Waste" containers will be in boxes (pipette tips). These should be disposed of in an autoclave bag and then placed in the MERI bin on the loading dock. "Liquid Waste" containers need to be placed in the sink with the addition of a little bit of bleach. Let the bleach and waste soak for at least 30 minutes, empty it out in the sink, rinse with distilled water (right-most faucet at the sink), and place it on the drying pads for washing through the glass washer machine.
Clean/tidy up the gel area. Empty waste containers of pipette tips into the sharps' containers (blue, "broken glass" cardboard box for non-biohazard tips). Refer to "Sharps and Laboratory Glass Disposal" poster for more information.
Parasite Tissue Storage. Any unused parasites should be frozen every week according to the tissue storage protocol listed in RNA Extraction. Stored tissue must be inventoried in the "Parasite Tissue and Nucleic Acid Inventory" spreadsheet and include a cryolabel with the Sample ID.
Glass washing and autoclaving. This is very important. Refer to Glass/Plastic Washing and Autoclave Operation for instructions.
Cleaning equipment. Wipe equipment in the lab with a Kimwipe lightly sprayed with 70% ethanol. This includes the heat block, microcentrifuges (outside and inside the machine), microscope glass stages, Qubit exterior, and thermal cycler exteriors. Be gentle and do not spray ethanol directly onto any electronics or equipment - remember these are very expensive machines and we want them to last a long time.
Refilling bleach/ethanol spray bottles. 10% bleach and 70% ethanol bottles need to be refilled. For bleach bottles, add 900 mL of distilled water (faucet with white handle) and 100 mL of Clorox bleach (located under the sink in Rm ??). For ethanol bottles, add 1000 mL of 70% ethanol. Check the bottles weekly to ensure all bottles are filled.
Pipette tip/consumables pipeline. Pipette tips and 1.5 mL tubes need to be autoclaved before use. Use the rack refill system and empty pipette tip boxes (in cabinet above plate reader in Rm 2961) to re-rack pipette tips. Place 1.5 mL tubes in a larger empty tip box. Autoclave for 60 min.
Preparing powder mixes for NGM and HGM plates. Measure out 1 L and 2 L amounts of dry ingredients for pouring NGM/HGM plates. Check to make sure all bottles have dry ingredients. Follow the Plate Pouring: NGM/HGM Plates recipe. Place bottles on the shelf in Rm 4210.
Plate pouring reagents and buffers. Cholesterol, potassium phosphate, calcium chloride, magnesium sulfate, etc. Follow instructions in C. elegans Recipes.
Pouring worm plates. This is an important part of our lab pipeline. We must always have enough plates poured, dried, seeded with bacteria, and ready for worms in order to execute our experiments. Refer to Plate Pouring: NGM/HGM Plates for instructions.
LB-AMP-Xgal-IPTG plates. Pour one sleeve of 10 cm plates following the recipe in General Lab Recipes. Place them in the 4°C in Rm 4206.
- X-gal stocks (50 mg/mL) recipe: Weigh 1 g of X-gal (MW: 408.64 g/mol) and dilute it in 20 mL of DMSO. Aliquot 1 mL into 1.5 mL tubes (makes 20).
- IPTG stocks (100 mM) recipe: Weigh 5 g IPTG (MW: 238.31 g/mol) and dilute it in 210 mL distilled water. Aliquot in 10 mL volumes into 15 mL conical tubes (makes 21).
Mosquito Carton Assembly Follow the instructions outlined in Mosquito Carton Assembly.
Agarose pads for injections. Make a large batch of agarose pads (20+) when running low. Follow the protocol in Microinjection.
Pull needles for mosquito injections Follow the instructions outlined in C. elegans Microinjection.

Equipment Maintenance (assigned)

(See Equipment Checklist)

Microinjection setup. See Injection Rig Preparation for maintenance instructions.
Water-jacketed incubators. The water-jacketed incubator has many critical maintenance requirements, including ensuring humidity pans are full, adding rust inhibitor to the water jacket, and completing full decontamination procedures. See Water-jacketed Incubators for full instructions. The water-jacketed incubator requires replacement of CO₂ tanks before tanks empty. See Connecting CO₂ Tanks for full instructions.
Freezer defrosting and scraping. Instructors for -80 and -20 C ... (?)
C. elegans incubators..** Wipe the interior of the 15/20°C C. elegans incubatorss with 70% ethanol once a month. Defrost 15C incubator 2-3 times per year.
Liquid N₂ tank. The liquid N₂ tank stores our critical backup copies of transgenic worm strains and transformed bacterial stocks. Follow Liquid N₂ Tank Maintenance for maintenance instructions.
Eye wash station. The eye wash station in Rm 2961 needs to be flushed o
Multidrop^TM Combi Reagent Dispenser. The dispensing cassettes must be recalibrated on a monthly basis. Refer to the Multidrop Dispenser Protocol for the calibration protocol.
AquaMax Plate Washer. Refer to the Cleaning protocols for instructions on the different types of cleaning (water and AquaMax cleaning solution).
High-context imager (ImageXpress). Communicating and managing vendor service requests to maintain the ImageXpress system in working condition. This includes virtual trouble-shooting with technical specialists and scheduling on-site service requests.