Difference between revisions of "20.109(S18):Prep notes for M3"
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'''after the laboratory:''' | '''after the laboratory:''' | ||
| − | * | + | *incubate for at least 24 hours, or for as long as needed given class schedule |
| − | ** | + | *add Fe2+ |
| + | **add 26.7 mL of 25 mM (NH<sub>4</sub>)2Fe(SO<sub>4</sub>)<sub>2</sub> added to 50 mL phage-AuNP, resulting in a final concentration of 8.7 mM | ||
| + | **stir for 12 hours, and for no more than ~14 hours | ||
*quench biomineralization | *quench biomineralization | ||
| − | **add 26.7 mL | + | **add 26.7 mL 25 mM Na:PO4 stock solution, resulting in final concentration of Fe and PO<sub>4</sub> of 6.45 mM |
| − | **stir for | + | ***(2.6 g monosodium phosphate monohydrate, 21.8 g disodium phosphate heptahydrate, up to 100 mL in dH<sub>2</sub>O) |
| + | **stir for 10 minutes | ||
*check pH | *check pH | ||
| − | **adjust to 6.5-7.5 with 50 mM | + | **adjust to 6.5-7.5 with 50 mM NaOH |
**stir for 6 hours | **stir for 6 hours | ||
===M3D2=== | ===M3D2=== | ||
'''prior to laboratory:''' | '''prior to laboratory:''' | ||
| − | * | + | *prepare aliquots of phage and Na:PO<sub>4</sub> for biomineralization demonstration |
'''per team:''' | '''per team:''' | ||
Latest revision as of 19:04, 17 April 2018
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:
- incubate for at least 24 hours, or for as long as needed given class schedule
- add Fe2+
- add 26.7 mL of 25 mM (NH4)2Fe(SO4)2 added to 50 mL phage-AuNP, resulting in a final concentration of 8.7 mM
- stir for 12 hours, and for no more than ~14 hours
- quench biomineralization
- add 26.7 mL 25 mM Na:PO4 stock solution, resulting in final concentration of Fe and PO4 of 6.45 mM
- (2.6 g monosodium phosphate monohydrate, 21.8 g disodium phosphate heptahydrate, up to 100 mL in dH2O)
- stir for 10 minutes
- add 26.7 mL 25 mM Na:PO4 stock solution, resulting in final concentration of Fe and PO4 of 6.45 mM
- check pH
- adjust to 6.5-7.5 with 50 mM NaOH
- stir for 6 hours
M3D2
prior to laboratory:
- prepare aliquots of phage and Na:PO4 for biomineralization demonstration
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
