BISC 219/F10:Media Recipes
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) |
| 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 PBS (phosphate buffered saline) | 137 mM NaCl,2.7 mM KCl,4.3 mM Na2HPO4,1.47 mM KH2PO4,Adjust to a final pH of 7.4. |
*NOTE: These concentrations are STOCK concentrations - not working concentrations!
Links to Labs& Project Info
Series1:
Worm Info
Lab 1: Worm Boot Camp & Sex-Linked or Autosomal Start
Lab 2: Sex-Linked or Autosomal Finale
Series2:
Background: Classical Forward Genetics and Gene Mapping
Lab 2: Mutant Hunt
Lab 3: Linkage Test Part 1
Lab 4: Linkage Test Part 2, Mapping and Complementation
Lab 5: Finish Complementation; Mapping Con't
Lab 6: DNA sequence analysis; Mapping Con't
Lab 7: Complete Mapping: Score
Series3:
Schedule of Reverse Genetics Project
RNAi General Information
Media Recipes
Lab 5: Picking your gene to RNAi
Lab 6: Cloning your gene of interest
Lab 7: Picking your transformant
Lab 8: Plasmid purification and transformation
Lab 9: Induction of bacteria for RNAi
Lab 10: Scoring your worms and RNA purification
Lab 11: RT PCR reactions
