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Team:GeorgiaTech/Agar Plates and Media Protocol
From 2013.igem.org
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*Add (35 g/L of LB [[Agar]]) or (25 g/L [[LB Broth|LB broth]] and 15 g/L [[Agar]]) to a flask with distilled water | *Add (35 g/L of LB [[Agar]]) or (25 g/L [[LB Broth|LB broth]] and 15 g/L [[Agar]]) to a flask with distilled water | ||
| - | *Dissolve and autoclave for | + | *Dissolve and autoclave for 1 hour on fluid cycle |
*Antibiotics should be added at the correct concentration and volume when the [[agar]] has cooled to around 50°C or cool enough for hands to be touching the exterior of the flask. | *Antibiotics should be added at the correct concentration and volume when the [[agar]] has cooled to around 50°C or cool enough for hands to be touching the exterior of the flask. | ||
Latest revision as of 20:14, 27 September 2013
- Dissolve and autoclave for 1 hour on fluid cycle
- Antibiotics should be added at the correct concentration and volume when the agar has cooled to around 50°C or cool enough for hands to be touching the exterior of the flask.
- Concentrations of antibiotics:
- Chloramphenicol 34 µg/µL - Blue
- Kanamycin 50 µg/µL - Green
- Ampicillin 100 µg/µL - Red
- For every mL of LB agar solution, add 1 µL of antibiotics.
- Since I was using 300 mL of water, I added .3 mL of kanamycin
- The plates are poured and cooled, then marked with a marker to signify the antibiotic.
- Agar stabs are from the same protocol, just poured into tubes and capped.
- Media solution is made from LB broth (25 g/L), autoclaved, and antibiotics added to it.
- From the toothpick used to create an agar stab, the toothpick is placed into the media solution to start the culture.
