HL-5 Media

Janet Smith | Boston Biomedical Research Institute

Yield: 1 liter

ingredients

10 g proteose peptone (Oxoid, catalog# LP0037)
5 g yeast extract (Oxoid, catalog# LP0021)
10 g glucose
7.9 ml 0.2 M Na₂HPO₄ (sodium phosphate, dibasic)
12.9 ml 0.2M KH₂PO₄ (potassium phosphate, monobasic)

preparation

Dissolve all components in ∼950 ml RT water. Adjust pH to 6.4-6.6 with concentrated HCl. Add slowly; typically only 4-8 drops are needed for 1 L media. Add water to 1L and dispense into glassware (we usually put 200 ml in 250 ml bottles.) Autoclave for 30 minutes and remove promptly. Store at RT.

notes

Adding Na₂HPO₄ and KH₂PO₄ from liquid stocks is convenient, but when scaling up it may be preferable to weigh out solid chemicals. Here are the solid amounts (per liter):

0.35 g Na₂HPO₄⋅7H₂0 or 0.224 g Na₂HPO₄ (anhydrous)
0.35 g KH₂PO₄ (anhydrous)