Minnesota/25 June 2008

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1. Steak LB + antibiotic plate:

a. From colonies whose plasmids we’ve sequenced, pick same colony and streak plates.Plates 6-14 were placed in the incubator @ 37C to regrow picked colonies for approximately 3 hours.

b. New LB plates with ampicillin were streaked with cells from the picked colonies for Biobricks 6-14. For each, a sterile pipette tip was dipped in the colony, and then used to streak a region of the new plate. A second sterile pipette tip was used to streak out of this region into an unstreaked section. Streaked plates were placed in incubator @ 37C to grow.

2. Begin 2mL glycerol stock culture:

a. The same colonies that were picked for the initial plasmid prep and the streaked plates directly above were again picked with a sterile pipette tip, and the tip was placed in 2mL LB broth + antibiotic (1uL/mL). All tubes were flamed immediately after opening and before closing and work was done near a flame to prevent contamination.

b. These were grown for approximately 18 hours @ 37C with shaking @ 220 rpm's.

3. Transformation results

4. 2mL cultures for plasmid miniprep

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