Following are recipes for all standard media prepared by the Media Preparation Facility.
3% Agar
- 7.5g Bacto Agar/ 250 ml H2O
4% Agar
- 10.0g Bacto Agar/ 250 ml H2O
40% Glucose
- 400 g/l Glucose
- autoclave
LB Broth, Miller
- 10.0 g/l Difco Bacto Tryptone
- 5.0 g/l Difco Yeast Extrct
- 10.0 g/l NaCl
- 10.0 ml/l 1.0 M Tris pH 7.5
- autoclave
2X LB Broth, Miller
- double above g/l
- autoclave
1.5% Agar/LB
- 6.0 g Bacto Agar/ 400 ml LB
PBS
- 8.0 g/l NaCl
- 0.2 g/l KCl
- 1.15 g/l Na2HPO4
- 0.2 g/l KH2PO4
- adjust pH to 7.2
- autoclave
2X S
- 13.4 g/l Difco Bacto Yeast Nitrogen Base w/o amino acids
- autoclave
2X S w/o ura, trp
- 13.4 g/l Difco Bacto Yeast Nitrogen Base w/o amino acids
- 4.0 g/l amino acid powder w/o ura, trp (prepared by Amon Lab)
- autoclave
2X S w/o his, leu
- 13.4 g/l Difco Bacto Yeast Nitrogen Base w/o amino acids
- 4.0 g/l amino acid powder w/o his, leu (prepared by Amon Lab)
Terrific Broth
Solution 1 : Prepared by the Media Preparation Facility
- 12.0 g/l Difco Bacto Tryptone
- 24.0 g/l Difco Bacto Yeast Extract or EMD Yeast Extract
- 4.0 ml/l Glycerol
- 900 ml H2O
- autoclave
Solution 2: Prepared by the individual labs
- 23.1 g/l KH2PO4
- 125.4 g/l K2HPO4
- autoclave and cool
- add 100 ml Solution 2 to 900 ml Solution 1
Versene
- 8.0 g/l NaCl
- 0.2 g/l KCl
- 1.15 g/l Na2HPO4
- 0.2 g/l KH2PO4
- 0.2 g/l Na-EDTA
- 0.015 g/l Phenol red
- adjust pH to 7.2
- autoclave
1.05X YEP
- 10.5 g/l EMD Yeast Extract or Difco Bacto Yeast Extract
- 21.1 g/l Difco Bacto Peptone
- autoclave
2.22X YEP
- 22.2 g/l EMD Yeast Extract or Difco Bacto Yeast Extract
- 44.4 g/l Difco Bacto Peptone
- autoclave
Trypsin/EDTA
- 0.20 g/l EDTA
- 8.00 g/l NaCl
- 0.40 g/l KCl
- 0.35 g/l NaHCO3
- 1.00 g/l Glucose
- 0.04 g/l Trypsin ( 428 units/ml) (Sigma # T-4665)
- 0.0075 g/l Phenol red
- adjust pH to 7.0
- sterile filter (0.2 um)
- Freeze immediately after filtering