From 2009.igem.org
High eff. EcoRI: RFP, constit. RFP
Thaw e. Coli till ice disappears
Pipette 100x into tube (X2)
Add 10x of plasmid to e. Coli
Place on ice for 30 min
Heat shock at 42C for 30 sec
Ice for 5 mins
Add 950x SOC
Place at 37C for 60 mins
Spin 5 mins
Pipette out SOC
Resuspend in 150x SOC
Spread 50x and 100x onto amp plates
More mini prep
Spin tubes #1-8 for 3 minutes
Combine tubes into #8
Add 300x TENS
Mix by inversion
Add 150x sodium acetate
Mix by inversion
Spin 2.5 mins
Transferred Sup to clean tube added 1mL of EtOH (Ethanol)
Spin for 5 mins
Washed pellet in 70% EtOH 300x
Spin 5 mins
pst = B#3
Spe and Xba = B#4
Resuspend 30x TE
Pst = B #3
Spe and ba = B#4
DIGEST
10x DNA
3x Buffer/B#3/B#4
15x dH2O
2x Eco RI/Pst/SPe/Xba
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