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

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