20.109(S18):Prep notes for M3
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M3D1
prior to laboratory:
- prepare LB (10 g tryptone, 5 g yeast extract, 10 g NaCl, up to 1 L in dH2O)
per team:
- 2 x 40 mL overnight cultures of M13-infected XL1-Blue cells
- 30 mL 20% PEG-8000/2.5 M NaCl
- 50 mL deionized water
during the laboratory:
- 50 mL conical tubes
- 25 mL pipets
- 2 mL eppendorf tubes
- quartz cuvettes
- 250 mL erlenmeyer flasks
- ice buckets
- stirbars
after the laboratory:
- Aliquot ~ 10 mL of students phage-Fe incubating mixture and transfer to 20 mL glass vial. This will be used for demonstration purposes for their next lab section
- 12-16 hours after phage-Fe incubation, add 1 mL of Na:PO4 solution to each groups flask (result is a 10 mM phosphate solution)
M3D2
prior to laboratory:
- 1 M Na:PO4, pH = 7.5 (2.6 g monosodium phosphate monohydrate, 21.8 g disodium phosphate heptahydrate, up to 100 mL in dH2O)
per team:
- 1 mL aliquots of student's phage in 2 mL tubes
- 25 μL 1 M Na:PO4, pH = 7.5
- 50 μL 100% ethanol
during the laboratory:
- 50 mL conical tubes
- 15 mL conical tubes
- tweezers
- old TEM grids, for practice
- new TEM grids, 2 per team
- sign-up sheet for location of samples in TEM case
M3D3
during the laboratory:
- flat stainless steel plates
- smooth stainless steel roller
- stainless steel spatulas
- mortar and pestle
- 3/8 puncher
- superP carbon powder
- Teflon 8A, binder
M3D4
per team:
- none
M3D5
per team:
- none