Team:Wageningen UR/Project/ProtocolsProj1

From 2011.igem.org

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'''LB'''
'''LB'''
-
Ingredients:
+
''Ingredients:''
10 g Bacto-tryptone
10 g Bacto-tryptone
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'''SOB'''
'''SOB'''
-
Ingredients
+
''Ingredients''
0.5% (w/v) yeast extract
0.5% (w/v) yeast extract
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20 mM MgSO4
20 mM MgSO4
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Per liter:
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''Per liter''
5 g yeast extract
5 g yeast extract
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'''SOC'''
'''SOC'''
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Ingredients
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''Ingredients''
SOB  
SOB  
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Materials
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''Materials''
Detergent-free, sterile glassware and plasticware
Detergent-free, sterile glassware and plasticware
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CCMB80 buffer
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''CCMB80 buffer''

Revision as of 17:16, 8 September 2011

Building a Synchronized Oscillatory System

Main Project

Protocols

Media

LB

Ingredients:

10 g Bacto-tryptone

5 g yeast extract

10 g NaCl


SOB

Ingredients

0.5% (w/v) yeast extract

2% (w/v) tryptone

10 mM NaCl

2.5 mM KCl

20 mM MgSO4

Per liter

5 g yeast extract

20 g tryptone

0.584 g NaCl

0.186 g KCl

2.4 g MgSO4


SOC

Ingredients

SOB

20 mM glucose


CCMB80 buffer for preparation of chemically competent cells


Materials

Detergent-free, sterile glassware and plasticware

Table-top OD600nm spectrophotometer

SOB


CCMB80 buffer


10 mM KOAc pH 7.0 (10 ml of a 1M stock/L)

80 mM CaCl2.2H2O (11.8 g/L)

20 mM MnCl2.4H2O (4.0 g/L)

10 mM MgCl2.6H2O (2.0 g/L)

10% glycerol (100 ml/L)

Adjust pH DOWN to 6.4 with 0.1N HCl if necessary

Adjusting pH up will precipitate manganese dioxide from Mn containing solutions

Filter sterilize and store at 4°C

Slight dark precipitate appears not to affect its function

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