BISC 219/F10:Media Recipes: Difference between revisions
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== | == Ingredients and Stock Concentrations for some of the reagents used in Series 3 == | ||
{| border="1" | {| border="1" | ||
|+ | |+ | ||
! | ! Reagent !! Composition | ||
|- | |- | ||
! Solution 1* | ! Solution 1* | ||
| Line 18: | Line 18: | ||
! Luria-Bertoni Medium (LB) | ! Luria-Bertoni Medium (LB) | ||
| 1% Tryptone <br> 0.5% Yeast Extract <br> 1% NaCl <br> (2% agar in plates) | | 1% Tryptone <br> 0.5% Yeast Extract <br> 1% NaCl <br> (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<br> 1.7% Bacto-agar <br> 0.25% Bacto-peptone <br> 0.1% (vol/vol)cholesterol | |||
|- | |- | ||
! Ampicillin* | ! Ampicillin* | ||
| Line 29: | Line 32: | ||
|- | |- | ||
! 1% agarose gel | ! 1% agarose gel | ||
|1% (w/v) agarose in 1X | |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) | ||
|- | |- | ||
! loading dye | ! 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 | |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 | ! 1x PBST (phosphate buffered saline with Tween20) | ||
|137 mM NaCl,2.7 mM KCl,4.3 mM Na2HPO4,1.47 mM KH2PO4,Adjust to a final pH of 7.4. | |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. | ||
|- | |- | ||
|} | |} | ||
'''*NOTE: These concentrations are STOCK concentrations - not working concentrations!''' | '''*NOTE: These concentrations are STOCK concentrations - not working concentrations!''' | ||
<div class=noprint> | |||
==Links to Labs& Project Info== | ==Links to Labs& Project Info== | ||
Series1:<BR> | Series1:<BR> | ||
| Line 65: | Line 68: | ||
[[BISC 219/F10: RNAi Lab 10 | Lab 10: Scoring your worms and RNA purification]]<br> | [[BISC 219/F10: RNAi Lab 10 | Lab 10: Scoring your worms and RNA purification]]<br> | ||
[[BISC 219/F10: RNAi Lab 11 | Lab 11: RT PCR reactions]]<br><br> | [[BISC 219/F10: RNAi Lab 11 | Lab 11: RT PCR reactions]]<br><br> | ||
</div> | |||
Latest revision as of 06:56, 18 May 2011
Ingredients and Stock Concentrations for some of the reagents used in Series 3
| Reagent | Composition |
|---|---|
| Solution 1* | 50 mM glucose 10 mM EDTA (Ethylenediaminetetraacetic acid) 25 mM Tris, pH 8 |
| Solution 2* | 0.2 M NaOH 1% SDS (sodium dodecyl sulfate) |
| 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 |
| Ampicillin* | Stock = 50 mg/ml in water use 1:1000 in liquid cultures 1:500 in agar |
| Tetracycline* | Stock = 12.5mg/ml in 50% EtOH use 1:1000 in liquid 1:1000 in agar |
| 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) |
| 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. |
*NOTE: These concentrations are STOCK concentrations - not working concentrations!
