Haynes Lab:Notebook/Synthetic Biology and Bioinformatics for Predictable Control of Therapeutic Gene2/2012/08/20

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Cell Culturing

Using the KAH126 insert with the biobrick BBa_J176062 Vector MV2 (mammalian) with birobrick BBa_J176122

DNA Prep

  1. 2 sample tubes (0.5mL)
  2. Add 10uL H20 and 0.5uL of DNA to (1) and 10uL H20 to (2)
  3. Need 30uL of cell per transformation (there is 200uL total in a stock tube) * Cell Line (DH5𝛼_turbo)
  4. 5 min on ice, label plates (Ex: Amp Dh6𝛼T (1) KAH-MV2 CMH 8/16/12)
  5. Add to plate, add bead
  6. Shake plate back and forth
  7. Dispose of beads
  8. Incubate for ~9 hrs
  • Note: Make sure to put tips in biohazard
  • Note: Go to ATCC Website and find the other two cell lines
  • Look for Lipofectamine LTX, McCoy's 5A tissue culture medium, and Opti-MEM

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