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Team:Frankfurt/Notebook
From 2012.igem.org
(→LBampicillin-Agar) |
(→Agar Plate) |
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Every cluture medium has to be autoclaved. | Every cluture medium has to be autoclaved. | ||
=Agar Plate= | =Agar Plate= | ||
| - | ===LB< | + | ===LB<sub>ampicillin</sub>-Agar=== |
Add 2 % agar to LB-medium. After autoclaving and cooling-down to 60 °C steril ampicillin is added. Plates were poured. | Add 2 % agar to LB-medium. After autoclaving and cooling-down to 60 °C steril ampicillin is added. Plates were poured. | ||
| - | |||
===SCD-Agar=== | ===SCD-Agar=== | ||
Add 2 % agar to SCD-medium. After autoclaving and cooling-down steril amino acid solution is added. Dependent on the respective selective medium Histidin (0.25 mM), Trypthophan (0.19 mM), Uracil (0.44 mM) or Leucin (0.35 mM) is added. | Add 2 % agar to SCD-medium. After autoclaving and cooling-down steril amino acid solution is added. Dependent on the respective selective medium Histidin (0.25 mM), Trypthophan (0.19 mM), Uracil (0.44 mM) or Leucin (0.35 mM) is added. | ||
===YEPD<sub>G418</sub>-Agar=== | ===YEPD<sub>G418</sub>-Agar=== | ||
Add 2 % agar to YEPD-medium. After autoclaving and cooling-down sterile G418 (final concentration 2g/l) is added. | Add 2 % agar to YEPD-medium. After autoclaving and cooling-down sterile G418 (final concentration 2g/l) is added. | ||
| + | |||
=Gel Electrophoresis= | =Gel Electrophoresis= | ||
{|class=wikitable float:right | {|class=wikitable float:right | ||
Revision as of 14:53, 14 September 2012
 | Home | Team | Project | Organisms | New Yeast RFC | Notebook | Registered Parts | Modeling | Safety | Attributions | Official Team Profile |
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You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well.
Contents |
Methods and Protocols
Culture Medium
| Full Medium (YEPD) for Yeast | |||
|---|---|---|---|
| Yeast Extract | 1 % (weight/volume) | ||
| Pepton | 2 % (w/v) | ||
| Glucose | 2 % (w/v) | ||
| Synthetic Complete Medium (SC) for Yeast | |||
|---|---|---|---|
| Yeast Nitrogen Base | 0.17 % (w/v) | ||
| Ammoniumsulfate | 0.5 % (w/v) | ||
| Amino Acid Mix* | 50 ml/l | ||
| Histidin** | 0.25 mM | ||
| Tryptophan** | 0.19 mM | ||
| Leucin** | 0.35 mM | ||
| Uracil** | 0.44 mM | ||
pH has to be regulated with KOH to pH=6.3
!* contains no His, Leu, Trp and Uracil
** addition of this components depents on the respective selection medium
| SOC-Medium for Regeneration of transformed Escherichia coli`s after Electroporation | |||
|---|---|---|---|
| Trypton | 2 % (w/v) | ||
| Yeast Extract | 0.5 % (w/v) | ||
| NaCl | 10 mM | ||
| KCl | 2,5 mM | ||
| MgCl2 | 10 mM | ||
| MgSO4 | 10 mM | ||
| Glucose | 20 mM | ||
pH has to be regulated to pH=6.8-7.0
| Full Medium (YEPD) for Yeast | |||
|---|---|---|---|
| Yeast Extract | 1 % (weight/volume) | ||
| Trypton | 0.5 % (w/v) | ||
| NaCl | 0.5 % (w/v) | ||
pH has to be regulated with NaOH to pH=7.5
Every cluture medium has to be autoclaved.
Agar Plate
LBampicillin-Agar
Add 2 % agar to LB-medium. After autoclaving and cooling-down to 60 °C steril ampicillin is added. Plates were poured.
SCD-Agar
Add 2 % agar to SCD-medium. After autoclaving and cooling-down steril amino acid solution is added. Dependent on the respective selective medium Histidin (0.25 mM), Trypthophan (0.19 mM), Uracil (0.44 mM) or Leucin (0.35 mM) is added.
YEPDG418-Agar
Add 2 % agar to YEPD-medium. After autoclaving and cooling-down sterile G418 (final concentration 2g/l) is added.
Gel Electrophoresis
| Agarose Gel (1x) | |||
|---|---|---|---|
| TAE puffer | 1x | ||
| Agarose | 1 % (w/v) | ||
Solve agarose in TAE by boiling it. After cooling-down to 55-60 °C gel is poured.
| TAE Puffer (50x) | |||
|---|---|---|---|
| EDTA | 18,6 g | ||
| Tris | 242g | ||
| Glacial Acetic Acid | 57,2 ml | ||
| Purified Water | 1000ml | ||
pH has to be regulated with glacial acetic acid to pH=8.
