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Last updated date: Mar 12, 2024 Views: 3692
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#2 Drop-out plate medium (1 Liter) 1.7 g Yeast nitrogen base without amino-acids without ammonium sulfate 5 g Ammonium sulfate 2 g drop-out powder mix 20g Bacto Agar 900 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. 20 g Glucose 100 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. After autoclaving mix the two solutions Stir slowly to avoid bubbles Let cool down on the side for 45 minutes, then pour 1L= 2 sleeves #3 Drop-out plate medium (1 Liter) 1.7 g Yeast nitrogen base without amino-acids without ammonium sulfate 5 g Ammonium sulfate 2 g drop-out powder mix 20g Bacto Agar 900 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. 20 g Raffinose 100 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. After autoclaving mix the two solutions Stir slowly to avoid bubbles Let cool down on the side for 45 minutes, then pour 1L= 2 sleeves #4 Drop-out plate medium (1 Liter) 1.7 g Yeast nitrogen base without amino-acids without ammonium sulfate 5 g Ammonium sulfate 2 g drop-out powder mix 20g Bacto Agar 900 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. 20 g Raffinose 20 g Galactose 100 mls MilliQ water Stir, but ingridients will not dissovle Autoclave at 121 C for 25 minutes. After autoclaving mix the two solutions Stir slowly to avoid bubbles Let cool down on the side for 45 minutes, then pour 1L= 2 sleeves YPD plate medium (1 Liter) 10 g yeast extract 20 g peptone 20g Bacto Agar 900 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. 20 g Glucose 100 mls MilliQ water Stir, but ingridients will not dissovle Autoclave at 121 C for 25 minutes. For plates with antibiotics add the following: 4 ml G418 (50 mg/ml) = Kan or Geniticin 300 ul NAT (200 mg/ml) = clonNAT 4 ml Hygromycin B (50 mg/ml) 1 ml Chloramphenicol (40 mg/ml) Let cool down on the side for 45 minutes Add antibiotic and stir slowly to avoid bubbles Pour plates right away 1L= 2 sleeves Kac – sporulation plate media (1 Liter) 20 g Potassium acetate 20 g Bacto Agar 100 mls MilliQ water Adjust to pH 7.0 Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. Let cool down on the side for 45 minutes, then pour 1L= 2 sleeves 5 FoA plate media (1 Liter) 3.35 g Yeast nitrogen base without amino-acids without ammonium sulfate 10 g glucose 1g drop-out powder mix (Complete) 1000 mls MilliQ water Adjust to pH 7.0 Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. Let cool down on the side for 45 minutes, then pour 1L= 2 sleeves #2 Drop-out liquid medium (1 Liter) 1.7 g Yeast nitrogen base without amino-acids without ammonium sulfate 5 g Ammonium sulfate 2 g drop-out powder mix 900 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. 20 g Glucose 100 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. After autoclaving mix the two solutions Stir slowly to avoid bubbles Let cool down on the side for 45 minutes, then pour 1L= 2 sleeves #3 Drop-out liquid medium (1 Liter) 1.7 g Yeast nitrogen base without amino-acids without ammonium sulfate 5 g Ammonium sulfate 2 g drop-out powder mix 900 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. 20 g Raffinose 100 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. After autoclaving mix the two solutions Stir slowly to avoid bubbles Let cool down on the side for 45 minutes, then pour 1L= 2 sleeves #4 Drop-out liquid medium (1 Liter) 1.7 g Yeast nitrogen base without amino-acids without ammonium sulfate 5 g Ammonium sulfate 2 g drop-out powder mix 900 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. 20 g Raffinose 20 g Galactose 100 mls MilliQ water Stir, but ingridients will not dissovle Autoclave at 121 C for 25 minutes. After autoclaving mix the two solutions Stir slowly to avoid bubbles Let cool down on the side for 45 minutes, then pour 1L= 2 sleeves YPD liquid medium (1 Liter) 10 g yeast extract 20 g peptone 20g Bacto Agar 900 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. 20 g Glucose 100 mls MilliQ water Stir, but ingridients will not dissovle Autoclave at 121 C for 25 minutes. LB plate medium (1 Liter) 10 g Tryptone 5 g Yeast Extract 10 g Sodium Chloride 15 g Bacto Agar 1000 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. For plates with antibiotics add the following, aliquots stored at -20 deg: 1 ml Carbenicillin (100 mg/ml) = Ampicilin 1 ml Kanamycin (50 mg/ml) 1 ml Chloramphenicol (40 mg/ml) Let cool down on the side for 45 minutes Add antibiotic and stir slowly to avoid bubbles Pour plates right away 1L= 2 sleeves LB liquid medium (1 Liter) 10 g Tryptone 5 g Yeast Extract 10 g Sodium Chloride 1000 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. TB liquid medium (1 Liter) 47.6 g Terrific Broth powder (RPI) 6 ml Glycerol 1000 mls MilliQ water Stir until all ingredients are dissolved Autoclave at 121 C for 25 minutes. Drop-out powder mix Adenine 0.5g Alanine 2g Arginine 2g Asparagine 2g Aspartic acid 2g Cysteine 2g Glutamic acid 2g Glutamine 2g Glycine 2g Histidine 2g Myo-inositol 2g Isoleucine 2g Leucine 4g Lysine 2g Methionine 2g para-Aminobenzoic acid 0.2g Phenylalanine 2g Proline 2g Serine 2g Threonine 2g Tryptophan 2g Tyrosine 2g Uracil 2g Valine 2g Weight all the powders except the one/ones that you do not need. Put the mix in a mortar and use the pestle to grind the powder. Put the mix in a clean plastic container and shake several times. References to order all the ingredients Amino Acids Reference Sigma Adenine A-2786 5g L-Alanine A-7469 25g L-Arginine A-8094 25g L-Asparagine A-7074 25g L-Aspartic acid A-7219 100g L-Cysteine C-7477 25g L-Glutamic acid G-8415 100g L-Glutamine G-8540 25g L-Glycine G-8790 100g L-Histidine H-6034 25g Myo-inositol I-7508 50g L-Isoleucine I-7403 25g L-Leucine L-8912 25g L-Lysine L-9037 25g L-Methionine M-5308 25g para-Aminobenzoic acid A-9878 5g L-Phenylalanine P-5482 25g L-Proline P-5607 25g L-Serine S-4311 25g L-Threonine T-8441 25g L-Tryptophan T-8941 25g L-Tyrosine T-8566 25g Uracil U-1128 25g L-Valine V-0513 25g D-glucose Sigma, ref G-8270 D-Raffinose RPI, R20500-1000.0 D-Galactose RPI, G33000-500.0 Ammonium Sulfate Sigma, ref A-2939 Bacto Agar Difco, ref 2140-10 Yeast nitrogen base w/o aa w/o ammonium sulfate Difco, ref 0335-15 Bacto Triptone BD, 211705 Bacto Peptone BD, DF0118-17-0 Terrific Broth RPI, T15000-5000.
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