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From 2012.igem.org

Biobricks

Methods:

Transformation of E. coli competent cells (from Inoue et al., 1990 'High efficiency transformation of Escherichia coli with plasmids', Gene, vol. 96, pp. 23-28) Experimental procedure:

• 1)Competent E. coli BL21(DE3) competent cells were obtained after growth and storage at -80C.
• 2)The tubes were defrosted in team mates hand. They were then put on ice
• 3)Add 1-10 µl of ligation mix to each tube. Incubated on ice for 5 min.
• 4)Tubes were heat shocked at 42C in water bath for 30 seconds and returned to ice for 2 min.
• 5) 200 µl of SOC media was added to each tube and incubated in the 37C shaker for 1 hour.
• 6) Each tube of cells had 5 µl spread onto an LB (Ampicillin) plate and another 50 µl onto a second LB (Ampicillin) plate, using aseptic technique.
• 7) The plates were then placed in an incubator at 37C for growth overnight