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  
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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  
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QSV 1 liter
 
QSV 1 liter
  
Plate Codes
+
'''Plate Codes'''
  
 
Line color Media Antibiotic
 
Line color Media Antibiotic
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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)
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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)
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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)