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Team:Grinnell/Recipes
From 2011.igem.org
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| + | <h1>Recipes</h1> | ||
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<h2><a name="LB"></a>L Broth</h2> | <h2><a name="LB"></a>L Broth</h2> | ||
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<li>2g Bacto peptone</li> | <li>2g Bacto peptone</li> | ||
<li>1g Yeast extract</li> | <li>1g Yeast extract</li> | ||
| - | <li>1mL 1M | + | <li>1mL 1M MgSO<sub>4</sub>⋅7H<sub>2</sub>O</li> |
| - | <li>1mL 0.5M | + | <li>1mL 0.5M CaCl<sub>2</sub></li></ul> |
| + | |||
| + | <h2><a name="M2G"></a>M2G Minimal Broth</h2> | ||
| + | <ol> | ||
| + | <li>For 1L of 20x M2 Salts:<ul> | ||
| + | <li>17.4g Na<sub>2</sub>HPO<sub>4</sub></li> | ||
| + | <li>10.6g KH<sub>2</sub>PO<sub>4</sub></li> | ||
| + | <li>10g NH<sub>4</sub>Cl</li> | ||
| + | </ul></li> | ||
| + | <li>Autoclave this sol'n.</li> | ||
| + | <li>To make M2G: | ||
| + | <ol> | ||
| + | <li>Dilute 20x M2 salts sol'n to 1x and autoclave.</li> | ||
| + | <li>Add the following sterile sol'ns: | ||
| + | <ul> | ||
| + | <li>1mL 0.5M MgSo<sub>4</sub></li> | ||
| + | <li>10mL 20% Glucose</li> | ||
| + | <li>1mL 10mM FeSO<sub>4</sub>, 10mM EDTA (Sigma #F-0518, kept at 4°C)</li> | ||
| + | <li>500μL 1M CaCl<sub>2</sub> (Note: add this last to avoid precipitation)</li> | ||
| + | </ul> | ||
| + | </li> | ||
| + | </ol> | ||
| + | </li> | ||
| + | </ol> | ||
<h2><a name="Agar"></a>Agar Plates</h2> | <h2><a name="Agar"></a>Agar Plates</h2> | ||
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<br/><b>Note</b>: for liquid cultures in PYE broth, use half the concentration that would be used for a PYE plate (LB remains the same for liquid and sold cultures). | <br/><b>Note</b>: for liquid cultures in PYE broth, use half the concentration that would be used for a PYE plate (LB remains the same for liquid and sold cultures). | ||
| + | |||
| + | <h2><a name="freezerStocks"></a>Freezer Stocks</h2> | ||
| + | To an appropriate tube add: | ||
| + | <ul> | ||
| + | <li>700μL overnight liquid culture</li> | ||
| + | <li>300μL 50% glycerol sol'n</li> | ||
| + | </ul> | ||
| + | Then place in storage at -80°C. | ||
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Latest revision as of 19:55, 28 September 2011
Recipes
L Broth
For 1L:- 10g Tryptone
- 5g Yeast extract
- 5g NaCl
PYE Broth
For 1L:- 2g Bacto peptone
- 1g Yeast extract
- 1mL 1M MgSO4⋅7H2O
- 1mL 0.5M CaCl2
M2G Minimal Broth
- For 1L of 20x M2 Salts:
- 17.4g Na2HPO4
- 10.6g KH2PO4
- 10g NH4Cl
- Autoclave this sol'n.
- To make M2G:
- Dilute 20x M2 salts sol'n to 1x and autoclave.
- Add the following sterile sol'ns:
- 1mL 0.5M MgSo4
- 10mL 20% Glucose
- 1mL 10mM FeSO4, 10mM EDTA (Sigma #F-0518, kept at 4°C)
- 500μL 1M CaCl2 (Note: add this last to avoid precipitation)
Agar Plates
Add 15g Bacto-agar to 1L of liquid media before autoclaving.Antibiotics Kitchen
| Antibiotic | Color Code | Stock (mg/mL) | LB Plate Conc (μg/mL) | PYE Plate Conc (μg/mL) |
|---|---|---|---|---|
| Ampicillin (amp) | Red | 50 | 100 | 50 |
| Carbenicillin (carb) | Red | 50 | 50 | 50 |
| Chloramphenicol (chlor) | 2 Orange | 5 (in EtOH) | 30 | 1 |
| Kanamycin (kan) | blue | 50 | 50 | 25 |
| Nalidixic acid (nal) | Green | 10 | 20 | 20 |
| Tetracycline (tet) | Black | 2 (in EtOH) | 12 | – |
| Oxytetracycline (oxytet) | Black | 2 (in EtOH) | − | 2 |
| Spectinomycin (spec) | Purple | 20 | 50 | 100 |
Note: for liquid cultures in PYE broth, use half the concentration that would be used for a PYE plate (LB remains the same for liquid and sold cultures).
Freezer Stocks
To an appropriate tube add:- 700μL overnight liquid culture
- 300μL 50% glycerol sol'n
