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
Hank's Balanced Salt Solution
Sigma # H-6136
- 0.01 g/l Na2HPO4
- 2.38 g/l Hepes
- 0.35 g/l NaHCO3
- adjust pH to 7.2
- sterile filter (0.2 um)
Hepes-Buffered Saline
- 7.07 g/l NaCl
- 0.40 g/l KCl
- 0.043 g/l Na2HPO4
- 0.060 g/l KH2PO4
- 1.00 g/l Glucose
- 4.77 g/l Hepes
- 0.0075 g/l Phenol red
- adjust pH to 7.3
- sterile filter (0.2 um)
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
10X PBS
- 80.0 g/l NaCl
- 2.0 g/l KCl
- 11.5 g/l Na2HO4
- 2.0 g/l KH2PO4
Note: neither pH adjusted nor sterilized
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)
2X S w/o 6 aa
- 13.4 g/l Difco Bacto Yeast Nitrogen Base w/o amino acids
- 4.0 g/l amino acid powder w/o ade, his, leu, lys, trp,ura (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
DME+/Hepes
SAFC Biosciences #56499C with: 4500 mg/l glucose, 110 mg/l Na pyruvate
- 0.116 g/l L-arginine*HCl
- 0.036 g/l L-asparagine
- 0.216 g/l L-glutamine
- 2.38 g/l Hepes
- 2.0 g/l NaHCO3
- Do NOT pH
- sterile filter (0.2 um)
DME-LG
Gibco # 31600 with 1000 mg/l glucose and 110 mg/l Na pyruvate
- 2.0 g/l NaHCO3
- adjust pH to 7.15
- sterile filter (0.2 um)
F-12 (Ham)
Gibco #21700
- 2.0 g/l NaHCO3
- adjust pH to 7.15
- sterile filter (0.2 um)
MEM alpha
Gibco #12000
- 2.2 g/l NaHCO3
- adjust pH to 7.15
- sterile filter (0.2 um)
MEM- (S-MEM, JOKLIKS)
Sigma # 56449C
- 2.0 g/l NaHCO3
- adjust pH to 7.2
- sterile filter (0.2 um)
MEM w/Erle's Salts
Gibco # 41500 with: Earle's salts (non-essential amino acids, L-glutamine)
- 1.5 g/l NaHCO3
- adjust pH to 7.2
- sterile filter (0.2 um)
RPMI
Gibco # 31800
- 0.30 g/l Glutamine
- 2.20 g/l NaHCO3
- adjust pH to 7.15
- sterile filter (0.2 um)
Pen/Strep
Mediatech #MT30-002-CI (VWR 45000-652)
10,000 IU/ml penicillin/10,000 ug/ml streptomycin
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