Reto's Buffers & Media Book
| Compound | 200 ml | ||
| Glycine | 75.07 | 1.5 g | |
| 1 M MgCl2 | 2 ml | ||
| KOH | |||
| H2O | to 200 ml |
| Compound | 50 ml | ||
| Acetic Acid 98% (~17 M) | 3 ml | ||
| H2O | to 50 ml |
Acrylamide/Bisacrylamide
| Compound | 500 ml | ||
| Acrylamide | 100 g | ||
| Bisacrylamide | 4 g | ||
| H2O | to 500 ml |
| Compound | 500 ml | ||
| Glucose | 4.5 g | ||
| Tris | 1.5 g | ||
| EDTA 0.25 M | 20 ml | ||
| H2O | to 450 ml | ||
| Adjust pH 8.0 with HCl | |||
| H2O, Autoclave, store at 4°C | to 500 ml |
| Compound | 100 ml | ||
| Lysozyme | 0.02 g | ||
| Alkaline Lysis Buffer A | to 100 ml |
| Compound | 50 ml | ||
| SDS 20 % | 2.5 ml | ||
| NaOH 10 M | 1 ml | ||
| H2O | to 50 ml |
| Compound | 50 ml | ||
| KAc | 14.7 g | ||
| Acetic Acid 100 % | 5.75 ml | ||
| H2O | to 50 ml |
| Compound | 100 ml | ||
| Amidoblack | 0.1 g | ||
| Acetic Acid 100 % | 7.5 ml | ||
| Methanol | 20 ml | ||
| H2O | to 100 ml |
| Constituent | ml stock for 1 liter medium | ||
| Adenine sulfate | |||
| Uracil | |||
| L-tryptophan | |||
| L-histidine-HCl | |||
| L-arginine-HCl | |||
| L-methionine | |||
| L-tyrosine | |||
| L-leucine | |||
| L-isoleucine | |||
| L-lysine-HCl | |||
| L-phenylalanine | |||
| L-glutamic acid | |||
| L-aspartic acid | |||
| L-valine | |||
| L-threonine | |||
| L-serine |
3-Aminophtalhydrazide 250 mM
| Compound | 10 ml | ||
| 3-Aminophtalhydrazide | 442 mg | ||
| H2O | to 10 ml |
| Compound | 50 ml | ||
| Ammoniummolybdate x 4H2O | 2.1 g | ||
| H2O | to 30 ml | ||
| HCl fuming 37 % (~12 M) | 16.7 ml | ||
| H2O | to 50 ml |
Ampicillin 20 mg/ml pH 8.0
| Compound | 15 ml | ||
| Ampicillin | 300 mg | ||
| H2O | to 10 ml | ||
| Adjust pH 8.0 with NaOH | |||
| H2O, Filter sterilize | to 15 ml |
Antibiotics
| Antibiotics | Resistance Mechanism | ||
| Ampicillin | Inhibits cell wall synthesis by inhibiting formation of the peptidoglycan crosslink | b-lactamase (286 aa) is secreted into the periplasm and cleaves lactam antibiotic | |
| Chloramphenicol | Inhibits protein synthesis by interacting with the 50S ribosomal subunit and inhibiting the peptidyltransferase reaction. | 216 aa protein cytoplasmic protein which acetylates chloramphenicol | |
| Kanamycin Neomycin | Inhibits translocation and elicits miscoding by binding to the 30S ribosomal subunit. | 256 aa aminoglycoside phosphotransferase phosphorylates kanamycin, neomycin, etc. | |
| Tetracycline | Inhibits protein synthesis by preventing binding of aminoacyl tRNA to the ribosome A site. | 428 aa membrane protein catalyzing efflux of tetracyclin, probably by a cation symport mechanism. |
APS
| Compound | 10 ml | ||
| Ammoniumpersulfate | 1 g | ||
| H2O, store at 4°C | to 10 ml |
Ascorbate 0.5 M pH 6.0
| Compound | 50 ml | ||
| L-Ascorbic acid | 4.4 g | ||
| H2O | to 22 ml | ||
| Adjust pH 6.0 with NaOH 1 M | |||
| H2O | to 50 ml |
Asolectin (fatty acid free) 50 mg/ml
| Compound | 50 ml | ||
| Asolectin (aceton treated) | 2.5 g | ||
| Diethylether, store at -20°C | to 50 ml |
ATP 200 mM
| Compound | 10 ml | ||
| Adenosine-5'-Triphosphate | 1.102 g | ||
| H2O | to 5 ml | ||
| Adjust pH 6.5 with NaOH 1 M | |||
| H2O, store at -20°C | to 10 ml |
ATP Regenerating System (ARS) 40x
| Compound | 1 ml | ||
| Phospoenolpyruvate (PEP) | 4.16 mg | ||
| Pyruvate Kinase (PK) 5 U/ml | 20 ml | ||
| H2O, store at 4°C | to 1 ml |
| Compound | 10 ml | ||
| Aprotinin 1.5 mg/ml | 3.24 ml | ||
| PepstatinA 1 mg/ml in DMSO | 1.3 ml | ||
| Leupeptin x 1/2H2SO4 10.5 mM | 37.5 ml | ||
| 4-Aminobenzamidine x 2HCl | 396 mg | ||
| H2O | to 10 ml |
Bis-Tris 1M pH7.0
| Compound | 100 ml | ||
| Bis-Tris | 20.9 g | ||
| H2O | to 50 ml | ||
| Adjust pH 7.0 with HCl | |||
| H2O | to 100 ml |
Brij/Doc pH 8.0
| Compound | 100 ml | ||
| EDTA 0.25 M | 0.4 ml | ||
| Tris 1 M pH 8.0 | 1 ml | ||
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