Hartwell's complete (HC) medium

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Contents

Stock solutions

10x HC dropout solution, per liter, autoclave

  • Methionine 200 mg
  • Tyrosine 600 mg
  • Isoleucine 800 mg
  • Phenylalanine 500 mg
  • Glutamic acid 1000 mg
  • Threonine 2000 mg
  • Aspartic acid 1000 mg
  • Valine 1500 mg
  • Serine 4000 mg
  • Arginine 200 mg


10x YNB, per liter, autoclave

  • Yeast nitrogen base (without amino acids or ammonium sulfate) 14.5 g
    • Note: this results in 15% less of the vitamin/salt mix than Difco YNB w/o amino acids.
  • Ammonium sulfate 50 g


  • Alternatively, Yeast nitrogen base (without amino acids) 57.1 g
  • Ammonium sulfate 7.4 g


1x amino acid solutions, separate, autoclave, per liter

  • Uracil 1 g (0.5 g in 500 ml)
  • Adenine 1 g (0.25 g in 250 ml)
  • Lysine 10 g (2.5 g in 250 ml)
  • Tryptophan 10 g (1.0 g in 100 ml)
    • keep in light blocking bottle
  • Leucine 20 g (2.0 g in 100 ml)
  • Histidine 10 g (1.0 g in 100 ml)


Final HC medium

  • Autoclave 20 g of agar in 619 ml water
  • Add 100 ml 20% sterile glucose solution
  • Add 100 ml 10x YNB solution
  • Add 100 ml 10x HC dropout solution

Optionally add

  • Adenine solution 20 ml
  • Lysine solution 12 ml
  • Leucine solution 4 ml
  • Histidine solution 2 ml
  • Tryptophan solution 8 ml
  • Uracil solution 35 ml


Bring the final volume to 1 liter

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