Question

1. to 10. Describe stepwise how would you prepare 1 L of a defined solid medium that requires vitamins B1 and B2 and penicillin. The recipe you found only has the final composition: Solidifying agent Salts 1.5% Agar 0.3% (22mM) Potassium Phosphate (KH2PO4) 0.6% (22mM) Sodium Phosphate (Na2HPO4) 0.5% (85mM) Sodium Chloride (NaCl) 0.1% Ammonium Chloride (NH4Cl) 2mM Magnesium Sulfate (MgSO) 0.1mM Calcium Chloride (CaCb) 0.4% Glucose (4.0 g/L) Carbon source 1 HM Vitamin B1 1 HM Vitamin B12 50 μg/mL penicillin 30 μg/mL kanamycin Thermolabile vitamins Thermolabile antibiotics Hint: You must prepare the vitamins and the antibiotics as concentrated stocks in water, then filter-sterilize, keep in a sterile container, and add to the medium after it cools down by adding a particular volume to one liter of medium (adding a few microliters will not affect the much larger volume of the medium). You have all the filters you need, syringes, autoclave, pipettes and pipette tips, balances, etc. Remember to always work under sterile conditions Example: Penicillin is required at a final volume of 50 μg/mL. A 10 mg/mL penicillin stock is made by dissolving 100 mg of penicillin to 10 mL of diH20 (deionized water). The penicillin solution is filter-sterilized and collected in a sterile 15-mL plastic tube. To calculate the volume to use, you may apply the proportion CiVi - C2V2, where C1-10 mg/mL, V, unknown, C2 = 50 μg/mL and V2 = 1 L. After converting L to mL and μg to mg: (50 μg·mL-1)(1 L) 10 mg mL1 5 X 10 2 mie )(103 ml.) 10 mg-mL1

Answer 1

S+oc stock2- 、 0.01 ml mol.al t· 135%. 3 891m24 2 l_AMメル 四,001mm×1000 mL こ0.01mL