Standard LB-agar plates.
- 1L glass Pyrex bottle, rinsed with ddH2O
- Electric pipette
- 25 mL pipette tips
- 40 petri dishes
- 25 g powdered LB broth, Miller
- 30 g agar
- Fill bottle with ~400 mL ddH2O.
- Add the components.
- Fill the rest of the bottle with ddH2O to 1L.
- Shake until components are completely dissolved.
- Autoclave 45 min.
- Invert the bottle 10 times to make sure all agar is dissolved homogeneously.
- Place in 65 °C water bath 1 hr.
- Optional: add antibiotics to desired final concentration. (Standard is 500 μL 100mg/mL ampicillin for Noireaux lab). Invert bottle ten times to mix antibiotics. Do not shake or in any other way introduce bubbles.
- Label petri dishes with type of antibiotic (e.g. 'C' for chloramphenicol). In Noireaux Lab, we do not label ampicillin plates, since they are our standard plates. Plates that do not have antibiotics should be labeled with '0' (put a slash through the zero).
- Aliquot by 25 mL in petri dishes with the electric pipette. Avoid pushing all the liquid through at the end of the tip, as this adds bubbles. If bubbles are introduced, removed them with the electric pipette. Work quickly, as the agar at the bottom will begin to solidify after ~20 min.
- Cool on bench overnight.
- Store at 4 °C upside down.
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
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- Anecdotal observations that might be of use to others can also be posted here.
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- Sean P Corum SC 16:36, 20 June 2012 (EDT):
or instead, discuss this protocol.