COMPLEX MEDIA
YPAD medium or Also Called YPD plus Adenine
Yeast Extract - Peptone - Dextrose plus Adenine medium
6.0 gm yeast extract (Difco)
12.0 gm Peptone (Difco)
12.0 gm Glucose
60 mg adenine hemisulphate
600 ml distilled water
Bacto-agar (Difco) 10 g (added for agar medium)
Sterilise by autoclaving at 121°C for
15 min.
We always use a 1 liter flask and make 600 mls of media because it gives exactly 20 plates (1 Sleeve of plates). In addition this volume of liquid is more easily handled when pouring after autoclaving.
SYNTHETIC COMPLETE DROP OUT(SC drop-out)MEDIUM
4.0 gm Difco Yeast Nitrogen Base (w/o amino
acids) 4.0 gm
12.0 gm glucose
0.50 gm Synthetic Complete Drop Out Mix
600 ml distilled water
10.0 gm Difco Bacto Agar (add for solidmedium)
Add the ingredients (except Agar) to the water and mix and adjust the pH to 5.6 with 10 N NaOH. (THIS STEP IS IMPORTANT FOR highest efficiency TRANSFORMATION)
Autoclave at 121°C for 15 min.
Synthetic Complete drop-out Medium Mix (SC drop-out)
Supplement Amount Note the hint about making this media below
2.0 gm Adenine
hemisulfate*
2.0 gm Arginine HCl
2.0 gm Histidine HCl*
2.0 gm Isoleucine
2.0 gm Leucine*
2.0 gm Lysine HCl
2.0 gm Methionine
3.0 gm Phenylalanine
6.0 gm Homoserine
3.0 gm Tryptophan*
2.0 gm Tyrosine
1.2 gm Uracil*
9.0 gm Valine
Omit the appropriate component(s), indicated in bold type, to prepare SC-Ade, SC-His, SC-Leu, SC-Trp, SC- Trp/Leu, and SC-Trp/Leu/His. Combine the ingredients in a clean 250 ml plastic bottle. Add three or four clean glass marbles and shake vigorously to mix. This quantity will be sufficient for approximately 100 (600 ml) batches of SC drop-out medium. Some recipes call for 20 amino acids in the SC mixture however most yeast strains do not need all of them. In addition some recipes do not use homoserine due to the expense, it can be replaced by the addition of serine and threonine (2 grams each).
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