IGEM:MIT/2006/Notebook/2006-10-30

From OpenWetWare

Jump to: navigation, search

Contents

Existing constructs

  • pSB1AT3-J45400 pLac.IAOGD
    • have uncut DNA and XP cut (?)
  • pSB1AC3-J45250 osmY.BGD
    • have uncut DNA (206.4ng/μL)
  • pSB1AT3-J45320 pLac.SAGD
    • have uncut DNA and ES cut (61 ng/μL)
  • pSB1AT3-J45120 pTet.BSMT
  • pSB1AK3-J45180 osmY.QPI.BSMT
    • have uncut and XP cut (69 ng/μL)
  • pSB1AC3-J45700 (45120+45320) pTet.BSMT.pLac.SAGD
    • have uncut DNA (260ng/μL)

To assemble

  • pSB1AC3-J45320.J45180
  • pSB1AK3-J45250.J45400
  • pSB1AK3-J45250.J45700
  • pSB1AK3-J45700.J45400

Smell tests

  • Use 4.4μL stock IA-OH per 10mL culture
  • Use 40μL 0.5M SA per 10mL culture

  1. Coculture pSB1AC3-J45250 and pSB1AT3-J45120 without salicylic acid and isoamyl alcohol
  2. Coculture pSB1AC3-J45250 and pSB1AT3-J45120 with salicylic acid and isoamyl alcohol
  3. Coculture pSB1AC3-J45250 and pSB1AK3-J45180 without salicylic acid and isoamyl alcohol
  4. Coculture pSB1AC3-J45250 and pSB1AK3-J45180 with salicylic acid and isoamyl alcohol
  5. Coculture pSB1AC3-J45250/pSB3K3-J45400 and pSB1AT3-J45700 without salicylic acid and isoamyl alcohol
  6. Coculture pSB1AC3-J45250/pSB3K3-J45400 and pSB1AT3-J45700 with salicylic acid and isoamyl alcohol
  7. Coculture pSB1AT3-J45200 and pSB1AT3-J45120 without salicylic acid and isoamyl alcohol
  8. Coculture pSB1AT3-J45200 and pSB1AT3-J45120 with salicylic acid and isoamyl alcohol
  9. Coculture pSB1AT3-J45200 and pSB1AK3-J45180 without salicylic acid and isoamyl alcohol
  10. Coculture pSB1AT3-J45200 and pSB1AK3-J45180 with salicylic acid and isoamyl alcohol

Done

  • pSB1AK3-J45180 transformed into Top10 cells, plated on amp. Also innoculated amp liquid culture directly with the transformation mix. -austin
  • grow up overnight cultures for miniprep of 180,250,320,400,700. -austin
  • Take care of growing up yeast constructs -samantha
  • Look at making the demo better -team
Personal tools