Goal: Grow large amounts of the pET28+CDS7 plasmid, grow up large amounts of the chlorophenicol biobrick backbone, restriction digest both and ligate the two together.

1. Streak both plasmids on a plate. grow O/N (Day 1)
2. Inoculate a few minicultures with colonies. grow O/N (Day 2)
3. Miniprep the minicultures (Day 3)
4. Verify plasmid using gel electrophoresis (Day 3)
5. Digest plasmid with restriction enzymes (Day 4)
6. Gel purify restriction digests (Day 5)
7. Ligate the purified products (Day 5)
8. Transform bacteria with ligation mixture. grow o/n (Day 6)
9. Inoculate miniculture of any successful transformants. grow o/n (Day 7)
10. Make glycerol stock (Day 8)
11. Miniprep/gel electrophoresis to verify (Day 9)

• Courtesy Dionne
• Note: o/n denotes overnight

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