BISC 219/F10:Media Recipes: Difference between revisions

Revision as of 09:58, 12 November 2010

Wellesley College BISC 219 Genetics

Home     People/Info     Lab Calendars/Weekly Planner     OWW Basics     Resources       Glossary
Series1:Sex-linked or Autosomal        Series2:Classical (Forward) Genetics        Series3: Reverse Genetics Using RNAi
       Worm Info     Assignments


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!

@@ Line 37: / Line 37: @@
 |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 PBS (phosphate buffered saline)
+! 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.
 |-
 |}