|Solvent||Stock Concentration to
|Bromo-chloro-indolyl-galactopyranoside||X-gal||20mg/ml||Dimethylformamide (DMF)||250x||Use DMF in a chemical hood|
|Isopropyl β-D-1-thiogalactopyranoside||IPTG||0.1M||H2O||1000x||Light sensitive. Molecular weight: 238.31 g/mol|
Glycerol for freezing aliquots
We use sterile 80% glycerol in water. We dilute this 1:3 (400ul 80% glycerol; 1200ul culture) for creating freezer stocks with a final glycerol concentration of 20%.
Used to create 1x PBS for dilution or washing cells of any species.
For 1 liter:
- NaCl 80g
- KCl 2g
- Na2HPO4 14.4g
- KH2PO4 2.4g
Stir salts with MilliQ water. This may take a significant time to dissolve!
Adjust pH with NaOH to 7.4. This will take a significant amount of NaOH; add in 200ul increments.
For Tris buffer at pH 8.0 (at 25 degrees)
- 4.44 g/L Tris HCl
- 2.65 g/L Tris Base
Do not adjust pH or check the pH with the MEE lab pH probe. This will destroy the probe!
- Calculate the amount of catalase needed; final concentration should be 30,000U / ml. Each batch may require a different number of milligrams of catalase.
- Use the precise balance of Karl's lab to measure out the catalase. Use a folded piece of weighing paper instead of a weighboat.
- Put catalase into a 15ml or 50ml plastic tube.
- Pipet in PBS, measuring out the volume using a serological pipet.
- Gently shake until dissolved.
- Sterilely filter solution using a syringe filter and place into a fresh, sterile plastic tube.
- Label top with contents and date; cover with foil to prevent exposure to light.
- Keep in refrigerator