BISC219/F12: Media Recipes: Difference between revisions
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Tucker Crum (talk | contribs) (New page: {{Template:BISC219/F12}} <div style="padding: 10px; width: 720px; border: 5px solid #7E354D;"> == Ingredients and Stock Concentrations for some of the reagents used in Series 3 == {| bord...) |
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! Reagent !! Composition | ! Reagent !! Composition | ||
|- | |- | ||
! Solution 1* | ! Miniprep Solution 1* | ||
| 50 mM glucose<br> 10 mM EDTA (Ethylenediaminetetraacetic acid) <br> 25 mM Tris, pH 8 | | 50 mM glucose<br> 10 mM EDTA (Ethylenediaminetetraacetic acid) <br> 25 mM Tris, pH 8 | ||
|- | |- | ||
! Solution 2* | ! Miniprep Solution 2* | ||
| 0.2 M NaOH <br> 1% SDS (sodium dodecyl sulfate) | | 0.2 M NaOH <br> 1% SDS (sodium dodecyl sulfate) | ||
|- | |- | ||
! Solution 3* | ! Miniprep Solution 3* | ||
| 2.7M Potassium Acetate <br> 6.6M Acetic Acid | | 2.7M Potassium Acetate <br> 6.6M Acetic Acid | ||
|- | |- | ||
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! Nematode Growth Medium Lite(NGM lite)- Available in powder form from US Biological Cat. #N1005 (http://www.usbio.net) | ! Nematode Growth Medium Lite(NGM lite)- Available in powder form from US Biological Cat. #N1005 (http://www.usbio.net) | ||
| 0.3% NaCl<br> 1.7% Bacto-agar <br> 0.25% Bacto-peptone <br> 0.1% (vol/vol)cholesterol | | 0.3% NaCl<br> 1.7% Bacto-agar <br> 0.25% Bacto-peptone <br> 0.1% (vol/vol)cholesterol | ||
|- | |||
! Chemotaxis media | |||
| 2% bactoagar <br> 5 mM potassium phosphate buffer, pH = 6.0 <br> 1 mM CaCl<sub>2</sub> <br> 1 mM MgSO<sub>4</sub> | |||
|- | |- | ||
! Ampicillin* | ! Ampicillin* | ||
| Stock = 50 mg/ml in water <br> use 1:1000 in liquid cultures <br> 1:500 in | | Stock = 50 mg/ml in water <br> use at 1:1000 in liquid cultures <br> use at 1:500 in LB amp plates | ||
|- | |||
|- | |||
! Tetracycline* | |||
| Stock = 12.5 mg/ml in water <br> use at 1:1000 in liquid cultures and in solid medium plates | |||
|- | |- | ||
!IPTG | !IPTG |
Latest revision as of 06:18, 26 October 2012
Ingredients and Stock Concentrations for some of the reagents used in Series 3
Reagent | Composition |
---|---|
Miniprep Solution 1* | 50 mM glucose 10 mM EDTA (Ethylenediaminetetraacetic acid) 25 mM Tris, pH 8 |
Miniprep Solution 2* | 0.2 M NaOH 1% SDS (sodium dodecyl sulfate) |
Miniprep Solution 3* | 2.7M Potassium Acetate 6.6M Acetic Acid |
Luria-Bertoni Medium (LB) | 1% Tryptone 0.5% Yeast Extract 1% NaCl (2% agar in plates) |
Nematode Growth Medium Lite(NGM lite)- Available in powder form from US Biological Cat. #N1005 (http://www.usbio.net) | 0.3% NaCl 1.7% Bacto-agar 0.25% Bacto-peptone 0.1% (vol/vol)cholesterol |
Chemotaxis media | 2% bactoagar 5 mM potassium phosphate buffer, pH = 6.0 1 mM CaCl2 1 mM MgSO4 |
Ampicillin* | Stock = 50 mg/ml in water use at 1:1000 in liquid cultures use at 1:500 in LB amp plates |
Tetracycline* | Stock = 12.5 mg/ml in water use at 1:1000 in liquid cultures and in solid medium plates |
IPTG | Stock = 0.5 M use 0.5 mM in cultures |
1% agarose gel | 1% (w/v) agarose in 1X TAE buffer. (50X TAE Stock Solution= 242 g/L Tris Base (MW=121.1); 57.1 mL/L Glacial Acetic Acid; 100 mL/L 0.5 M EDTA; Add ddH2O to 1 Liter and adjust pH to 8.5 using KOH) |
DNA loading dye | 0.4 % Ficoll 400, 1.8 mM EDTA, 0.55 mM Tris-HCl, 0.00117 % SDS, 0.025 % Bromophenol Blue pH 8.0 @ 25°C |
1x PBST (phosphate buffered saline with Tween20) | 137 mM NaCl,2.7 mM KCl,4.3 mM Na2HPO4,1.47 mM KH2PO4, 0.25% -Adjust to a final pH of 7.4. |
50X TAE (use at 1X) | 2M Tris Base, 0.05M EDTA, 5.7% Glacial Acetic Acid, adjust to a final pH of 8.5 with KOH |
*NOTE: These concentrations are STOCK concentrations - not effective concentrations!