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Team:NTNU Trondheim/Protocols
From 2011.igem.org
(Difference between revisions)
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*10 mL 250 mM KCl (25mM) → 0,186 g / 10mL H20 | *10 mL 250 mM KCl (25mM) → 0,186 g / 10mL H20 | ||
*5 mL 2M MgCl2 (10mM) → 0,952 g / 5mL | *5 mL 2M MgCl2 (10mM) → 0,952 g / 5mL | ||
| + | |||
| + | Autoclave, cool, then add: | ||
| + | *20 mL 1M sterile-filtered glucose (20mM) | ||
| + | |||
| + | ==Ampicillin-plates:== | ||
| + | Use stock 200 mg/mL. → 100µg/mL in liquid agar. | ||
| + | I.E. 250 µL stock in 500 mL agar. Add when cool enough to pour. | ||
| + | |||
| + | ==Spectinomycin-plates:== | ||
| + | Use stock 100 mg/mL. → 100µg/mL in liquid agar. | ||
| + | I.E. 500 µL stock in 500 mL agar. Add when cool enough to pour. | ||
| + | |||
| + | ==Ampicillin-IPTG plates:== | ||
| + | Ampicillin 100 µg / mL (500 µL amp stock (200 mg/mL) in 500 mL agar) | ||
| + | IPTG 0,1 mM ( 63,5 µL IPTG stock (0,8 M)) | ||
Revision as of 07:51, 5 July 2011
Protocols / Recipes used in the lab
LB Broth
- Bacto-tryptone: 10 g/L
- Yeast extract: 5 g/L
- NaCl 5 g/L
Adjust pH to 7,4 with NaOH, autoclave.
LA
- Dissolve LB
- Add agar, 15 g/L
Adjust pH to 7,4 with NaOH, autoclave.
SOC:
- Bacto-tryptone: 20 g/L
- Yeast extract: 5 g/L
- NaCl 0,5 g/L
- 10 mL 250 mM KCl (25mM) → 0,186 g / 10mL H20
- 5 mL 2M MgCl2 (10mM) → 0,952 g / 5mL
Autoclave, cool, then add:
- 20 mL 1M sterile-filtered glucose (20mM)
Ampicillin-plates:
Use stock 200 mg/mL. → 100µg/mL in liquid agar. I.E. 250 µL stock in 500 mL agar. Add when cool enough to pour.
Spectinomycin-plates:
Use stock 100 mg/mL. → 100µg/mL in liquid agar. I.E. 500 µL stock in 500 mL agar. Add when cool enough to pour.
Ampicillin-IPTG plates:
Ampicillin 100 µg / mL (500 µL amp stock (200 mg/mL) in 500 mL agar) IPTG 0,1 mM ( 63,5 µL IPTG stock (0,8 M))
