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Team:Aachen/TestBench2
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Revision as of 21:31, 17 September 2014 by
Mosthege
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Help
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2D Biosensor
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Contents
1
Media
1.1
LB medium
1.2
Hartmans minimal medium (HM)
2
Agar Chips
3
Transformation
3.1
Heat Shock
3.2
Electroporation
4
PCR
4.1
Colony PCR
4.2
QuikChange
5
Clonings
5.1
Restriction Digest
5.2
Ligation
5.3
Gibson Assembly
Media
LB medium
weight components
5 g/L NaCl
10 g/L tryptone
5 g/L yeast extract
(15 g/L agar for plates)
fill up to 1 L with deionized water
mix well
by shaking
autoclave
autoclaving tape, caps slightly unscrewed
base of the pot has to be covered with deionized water
close lid
heat
level 3 until the pressure valve opens
reduce
heat level to 1.5
set timer to
20 minutes
turn heater off
wait until the pressure valve retracts
(30-45 minutes)
open, close caps & shake
for plates, wait until you can touch the bottle (
<60 °C
, clean bench!)
add antibiotics and
shake
(gloves!)
Hartmans minimal medium (HM)
Agar Chips
Transformation
Heat Shock
thaw cells on ice
add 1 µl of plasmid DNA
incubate on ice for 30'
heat shock at 42 °C for 60
incubate on ice for 5'
add 200 µl of SOC media
incubate at 37 °C for 2 h
plate 20 and 200 µl on plates with the appropiate antibiotic
Electroporation
PCR
Colony PCR
QuikChange
Clonings
Restriction Digest
Ligation
Gibson Assembly