Difference between revisions of "Bacterial Media"
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| − | 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 | ||
| + | |||
10 g Tryptone | 10 g Tryptone | ||
| + | |||
5 g NaCl | 5 g NaCl | ||
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 | ||
| + | |||
10 g Tryptone | 10 g Tryptone | ||
| + | |||
5 g NaCl | 5 g NaCl | ||
| + | |||
15 g BactoAgar | 15 g BactoAgar | ||
QSV 1 liter | QSV 1 liter | ||
| − | Plate Codes | + | '''SOC Broth''' |
| + | |||
| + | 2 g Bacto Tryptone | ||
| + | |||
| + | 0.5 g Bacto Yeast Extract | ||
| + | |||
| + | 0.2 mL 5M NaCl | ||
| + | |||
| + | 0.25 mL 1M KCl | ||
| + | |||
| + | 1 mL 1M MgCl2 | ||
| + | |||
| + | 1 mL 1M MgSO4 | ||
| + | |||
| + | 2 mL 1M Dextrose (Glucose) | ||
| + | |||
| + | 90 mL of dH2O | ||
| + | |||
| + | Mix, then fill up to 100 mL with dH2O | ||
| + | |||
| + | '''Plate Codes''' | ||
| − | + | #Black line only LB none | |
| − | Black only LB none | + | #Black line, Red line LB Amp |
| − | Black, Red | + | #Black line, Blue line LB Tet |
| − | Black, Blue LB Tet | + | #Black line, Green line LB Kan |
| − | Black, Green LB Kan | + | #Black line, Cyan line LB Chlor |
| − | Black, Cyan | ||
| − | 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 61: | ||
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) | ||
| + | |||
Dissolve in 50 ml 70% Ethanol | Dissolve in 50 ml 70% Ethanol | ||
| + | |||
Mix/vortex vigorously | Mix/vortex vigorously | ||
| + | |||
| + | Filter sterilize | ||
| + | |||
| + | Make 4 ml aliquot in foil covered 15 ml tubes | ||
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 | + | Light Sensitive! Keep plates in darkness |
| + | |||
| + | '''Kanamycin''' | ||
| + | Stock – 10 mg/ml | ||
Weigh out 0.5 g Kanamycin sulfate (Sigma K1377) | Weigh out 0.5 g Kanamycin sulfate (Sigma K1377) | ||
| Line 51: | Line 86: | ||
Working concentration 20 ug/ml (2 ml stock solution per liter) | Working concentration 20 ug/ml (2 ml stock solution per liter) | ||
| − | Chloramphenicol Stock – | + | '''Chloramphenicol''' |
| + | Stock – 35 mg/ml | ||
| − | Weigh out 1. | + | Weigh out 1.75 g Chloramphenicol (Sigma C0378) |
Add 50 ml 100% Ethanol | Add 50 ml 100% Ethanol | ||
Mix/vortex vigorously | Mix/vortex vigorously | ||
| − | Working concentration | + | Working concentration 35 ug/ml (1 ml stock solution per liter) |
| + | |||
| + | '''L-Arabinose''' | ||
| + | Stock – 250 mg/ml | ||
| + | |||
| + | Weigh out 2.5 g L-Arabinose | ||
| + | Add 10 ml dH2O | ||
| + | Mix/vortex vigorously | ||
| + | |||
| + | Working concentration 0.5 mg/ml (2 ml stock solution per liter) | ||
Latest revision as of 18:49, 3 July 2014
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
SOC Broth
2 g Bacto Tryptone
0.5 g Bacto Yeast Extract
0.2 mL 5M NaCl
0.25 mL 1M KCl
1 mL 1M MgCl2
1 mL 1M MgSO4
2 mL 1M Dextrose (Glucose)
90 mL of dH2O
Mix, then fill up to 100 mL with dH2O
Plate Codes
- Black line only LB none
- Black line, Red line LB Amp
- Black line, Blue line LB Tet
- Black line, Green line LB Kan
- Black line, Cyan line 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
Filter sterilize
Make 4 ml aliquot in foil covered 15 ml tubes
Working concentration 20 ug/ml (4 ml stock solution per liter)
Light Sensitive! Keep plates in darkness
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 – 35 mg/ml
Weigh out 1.75 g Chloramphenicol (Sigma C0378) Add 50 ml 100% Ethanol Mix/vortex vigorously
Working concentration 35 ug/ml (1 ml stock solution per liter)
L-Arabinose
Stock – 250 mg/ml
Weigh out 2.5 g L-Arabinose Add 10 ml dH2O Mix/vortex vigorously
Working concentration 0.5 mg/ml (2 ml stock solution per liter)
