Staff Maintenance Tasks

General Lab Upkeep

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 223 on the fridge when a duty 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.

  • 100 bp and 1000 bp ladders (from vectors). See Ladder Creation for instructions.

  • Loading dye. Make aliquots of 6x orange loading dye into 1.5 mL tubes. See the loading dye recipe in General Recipes.

  • dNTP stocks preparation. 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.

  • 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.

  • Agarose pads for injections. Make a large batch of agarose pads (20+) when running low. Follow the protocol in Microinjection.

  • Worm 2x lysis buffer. This needs to be made rarely, in large batches and frozen. See C. elegans Recipes.

  • M9 stock replenishment. Make three 1 L stocks of M9 in glass bottles when running low. Follow the recipe in C. elegans Recipes. Be sure to autoclave and then add 1 M MgSO4 once cool.

Equipment Maintenance

(See Equipment Checklist)

  • Injection setup maintenance. See Injection Rig Preparation for maintenance instructions.

  • Incubator maintenance. 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.

  • Replacing incubator CO2 tanks. The water-jacketed incubator requires replacement of CO2 tanks before tanks empty. See Connecting CO2 Tanks for full instructions.

  • Liquid N2 tank upkeep. 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.

Task Assignments

Task Member Responsible
50x TAE and 1x TAE refill KD
100 bp and 1000 bp ladder reaction NW
Loading dye stocks KD
dNTP preparation KD/ZH
M9 stock refill ZH
SOC Media KV
Making competent cells (DH5alpha) KV
Injection setup and needle puller maintenance KV
Incubator maintenance and CO2 tank replacement KV
Water-jacket incubator maintenance KV
Worm 2x lysis buffer KV
Agarose pads KV
Liquid N2 tank monitoring KV