Difference between revisions of "Bacterial Media"
Line 1: | Line 1: | ||
− | Missouri Western Synthetic Biology Protocols | + | '''Missouri Western Synthetic Biology Protocols |
− | Bacterial Media Prep | + | Bacterial Media Prep''' |
− | LB Broth | + | '''LB Broth''' |
5 g Yeast Extract | 5 g Yeast Extract | ||
Line 10: | Line 10: | ||
QSV 1 liter | QSV 1 liter | ||
− | LB Plates (makes about 35 plates) | + | '''LB Plates''' (makes about 35 plates) |
5 g Yeast Extract | 5 g Yeast Extract | ||
Line 19: | Line 19: | ||
QSV 1 liter | QSV 1 liter | ||
− | Plate Codes | + | '''Plate Codes''' |
Line color Media Antibiotic | Line color Media Antibiotic | ||
Line 28: | Line 28: | ||
Black, Cyan LB Chlor | Black, Cyan LB Chlor | ||
− | Ampicillin Stock – 50 mg/ml | + | '''Ampicillin''' |
+ | Stock – 50 mg/ml | ||
Weigh out 2.5 g Ampicillin, sodium salt (Sigma A0166) | Weigh out 2.5 g Ampicillin, sodium salt (Sigma A0166) | ||
Line 36: | Line 37: | ||
Working concentration 50 ug/ml (1 ml stock solution per liter) | Working concentration 50 ug/ml (1 ml stock solution per liter) | ||
− | Tetracycline Stock – 5 mg/ml | + | '''Tetracycline''' |
+ | Stock – 5 mg/ml | ||
Weigh out 0.25 g Tetracycline HCl (Sigma T7660) | Weigh out 0.25 g Tetracycline HCl (Sigma T7660) | ||
Line 44: | Line 46: | ||
Working concentration 20 ug/ml (4 ml stock solution per liter) | Working concentration 20 ug/ml (4 ml stock solution per liter) | ||
− | Kanamycin Stock – 10 mg/ml | + | '''Kanamycin''' |
+ | Stock – 10 mg/ml | ||
Weigh out 0.5 g Kanamycin sulfate (Sigma K1377) | Weigh out 0.5 g Kanamycin sulfate (Sigma K1377) | ||
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Working concentration 20 ug/ml (2 ml stock solution per liter) | Working concentration 20 ug/ml (2 ml stock solution per liter) | ||
− | Chloramphenicol Stock – 25 mg/ml | + | '''Chloramphenicol''' |
+ | Stock – 25 mg/ml | ||
Weigh out 1.25 g Chloramphenicol (Sigma C0378) | Weigh out 1.25 g Chloramphenicol (Sigma C0378) |
Revision as of 14:31, 31 May 2010
Missouri Western Synthetic Biology Protocols Bacterial Media Prep
LB Broth
5 g Yeast Extract 10 g Tryptone 5 g NaCl
QSV 1 liter
LB Plates (makes about 35 plates)
5 g Yeast Extract 10 g Tryptone 5 g NaCl 15 g BactoAgar
QSV 1 liter
Plate Codes
Line color Media Antibiotic Black only LB none Black, Red LB Amp Black, Blue LB Tet Black, Green LB Kan Black, Cyan LB Chlor
Ampicillin
Stock – 50 mg/ml
Weigh out 2.5 g Ampicillin, sodium salt (Sigma A0166) Dissolve in 25 ml dH20 Add 25 ml Ethanol
Working concentration 50 ug/ml (1 ml stock solution per liter)
Tetracycline
Stock – 5 mg/ml
Weigh out 0.25 g Tetracycline HCl (Sigma T7660) Dissolve in 50 ml 70% Ethanol Mix/vortex vigorously
Working concentration 20 ug/ml (4 ml stock solution per liter)
Kanamycin
Stock – 10 mg/ml
Weigh out 0.5 g Kanamycin sulfate (Sigma K1377) Dissolve in 50 ml dH20
Working concentration 20 ug/ml (2 ml stock solution per liter)
Chloramphenicol
Stock – 25 mg/ml
Weigh out 1.25 g Chloramphenicol (Sigma C0378) Add 50 ml 100% Ethanol Mix/vortex vigorously
Working concentration 25 ug/ml (1 ml stock solution per liter)