Stock Center Media

Contributed by Jakob Franke, December 2005
Download word file.

[INDEX]

HL5
Maltose HL5
MES-HL5
2 x HBS
SM-plates
SM/5 plates
LP agar
LP/2 agar
Sor
SorC
Standard Solution (SS) (Bonner's salts)

• HL5 (1 liter)
  Start with deionized water in a plastic beaker, and add reagents while stirring.
  • 10 g Dextrose (D-glucose)
  • 5 g BBL yeast extract
  • 5 g Proteose peptone (Difco)
  • 5 g Thiotone peptone (BBL)
  • 0.67 g Na₂HPO₄.7H₂O
  • 0.34 g KH₂PO₄
  • 0.05 g Dihydrostreptomycin-sulfate*

The pH should be 6.5 ± 0.1.
Add 25 ml to 125 ml erlenmeyer flasks. Cover with cotton plugs and aluminum foil.
Autoclave 20 minutes.
Store in the cold room, and dark.
* Dihydrostreptomycin-sulfate is stored in the refrigerator.
Warm up to room temperature before opening the bottle!

• Maltose HL5 (1 liter)
  • 20 g Maltose
  • 5 g yeast extract (BBL)
  • 5 g Proteose peptone (Difco)
  • 5 g Thiotone peptone (BBL)
  • 0.67 g Na₂HPO₄.7H₂O
  • 0.34 g KH₂PO₄
  • 0.05 g Dihydrostreptomycin-sulfate

pH should be 6.4-6.5.

• MES-HL5 (1 liter)
  • 10 g Dextrose (glucose)
  • 5 g Yeast extract (BBL)
  • 5 g Proteose peptone (Difco)
  • 5 g Thiotone peptone (BBL)
  • 1.3 g MES
  • 0.05 g Dihydrostreptomycin-sulfate

Adjust the pH to 7.1 with NaOH. It's important!
Autoclave 20 minutes.

• 2 x HBS (250 ml)
  • 4 g NaCl
  • 0.18 g KCl
  • 0.05 g Na₂HPO₄ (or 0.094 g Na₂HPO₄.7H₂O)
  • 0.5 g Dextrose
  • 2.5 g HEPES

Adjust pH to 7.0-7.1 with NaOH. It's important!
Filter sterilize.
Keep frozen in 50 ml aliquots.

• SM-plates (1 liter) Start with 1 L dH₂O, and weigh out:
  • 10 g Glucose (dextrose)
  • 10 g Bacto peptone (Difco)
  • 1 g Bacto yeast extract
  • 0.5 g MgSO₄ (or 1 g MgSO₄.7H₂O)
  • 1.8 g KH₂PO₄ (13.2 mM)
  • 1.2 g K₂HPO₄ (6.9 mM)

The pH should be 6.4-6.5. If too low, adjust with 1 N KOH. If too high, adjust with 1 N HCl.
Add 15 g agar (Spectrum brand).
Autoclave 30 minutes.
Cool about 30 minutes.
Pour plates (30-35 plates per L).

• SM/5 plates (1 liter) Start with 1 L dH₂O, and weigh out:
  • 2 g Glucose (dextrose)
  • 2 g Bacto peptone (Difco)
  • 0.2 g Bacto yeast extract
  • 0.1 g MgSO₄ (or 0.2 g MgSO₄.7H₂O)
  • 1.9 g KH₂PO₄
  • 1.0 g K₂HPO₄

Check pH: should be 6.5 ± 0.1.
Add 15 g agar (Spectrum brand).
Autoclave 30 minutes.
Pour 30-35 plates per liter.

• LP agar (also called 0.1LP agar)
  • 1 g Lactose
  • 1 g Bacto peptone (Difco)
  • 15 g Agar (Spectrum brand)
  • 1 L Deionized water

Autoclave 20 minutes.

• LP/2 agar (1 liter) (also called 0.1LP/2 agar)
  • 0.5 g Lactose
  • 0.5 g Bacto peptone (Difco)
  • 15 g Agar (Spectrum brand)
  • 1 L Deionized water

Autoclave 20 minutes.

• Sor (Sörensen's buffer) (1/60 M phosphate buffer, pH 6; PMID: 4384821; d2776)
  • 4 L dH₂O
  • 8.0 g KH₂PO₄
  • 1.16 g Na₂HPO₄ (or 2.2 g Na₂HPO₄.7H₂O)

pH should be 6.0 ± 0.1

• SorC (Sor with 50 µM Ca++)
  • Add 4 ml 50 mM CaCl₂ (or 0.2 ml 1 M CaCl₂) to 4 L Sor, and autoclave.

• Standard Solution (SS) (also called Bonner's salt) (1 liter) (d0096)
  • 1 L deionized water
  • 0.6 g NaCl
  • 0.75 g KCl
  • 0.3 g CaCl₂ (or 0.4 g CaCl₂.2H₂O)

Autoclave.

[INDEX]