Bacto-agar medium

YT-Cm25 plates 2YT-Cm25 medium supplemented with 1.5% (w/v) bacto agar. 

Prepare a basal layer of 7 ml of medium (Eagle s Glasgow modification containing tryptose and calf serum) containing 0.5% Difco Bacto agar in a 6 cm dish. 

The mouse L-929 fibroblast line was cultivated in Eagle s Minimal Essential Medium (MEM) plus 10% calf serum. Cells were seeded in 100-mm-diameter cell culture plates at 4 x 106 cells per plate and allowed to become established for 24 hr prior to use. After the monolayer was washed, 10 mL of an agar overlay consisting of 2% Bacto-Agar and 2 x MEM was added to each plate and allowed to solidify. 

LB agar (LB medium +15 g/L bacto agar, mix and autoclave to dissolve), and plates filled with LB agar containing 50 pg/ml. ampicillin. Ampicillin is added before plating but after cooling to 40°C. 

To maintain the cells, they are grown on slants containing the peptone medium plus 1.5% Bacto-Agar. 

Ampicilhn 50 mg/mL in water, sterilize by filtering through a 0.22-pm filter 4. LB-ampicillin agar plates Add 1.4 g of bacto-agar to 100 mL of LB medium,... 

Luria-Bertani (LB) medium sodium chloride (NaCl, 10 g/L), Bacto tryptone (10 g/L), and Bacto yeast extract (5 g/L). Add 15 g/L Bacto agar to LB medium to prepare LB agar for plating. Dissolve in ddH O and autoclave. 

EMM complete solid medium Dissolve 12.33 g of EMM without dextrose and 30 g of glucose in 470 mL of distilled H2O. Add 150 mg of leucine and 75 mg each of adenine, histidine, lysine, and uracil (for selective media, do not add the supplement for which selection is based). Dissolve by mixing under low heat and filter-sterilize. Combine with 490 mL of water plus 20 g of Bacto agar that has been autoclaved in a 2 L flask. 

Materials for media (Bacto Tryptone, Bacto Yeast Extract and Difco SOB Medium, agar powder and NaCl) and antibiotics (ampicillin sodium and kanamycin sulfate). 

LB-IPTG-Xgal plates LB medium + 15 g/L agar (either technical or Bacto grade). Autoclave, cool to <70 °C, add 1 mL IPTG/Xgal and pour into petri plates. Store plates at 4 °C in the dark. 

Assay Medium. Herrell el al. (16) recommend the use of fluid thioglycol-ate broth with the following composition yeast extract 5 g., Bacto-casitone 15 g., glucose 5 g., sodium chloride 2.5 g., Z-cystine 0.75 g., thioglycollic acid 0.3 ml., agar 0.75 g., resazurin (certified) 0.001 g., and distilled water 1 liter the final pH of this medium should be 7.1. 

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