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/&mu;L)
 * pSB1AT3-J45320 pLac.SAGD
 * have uncut DNA and ES cut (61 ng/&mu;L)
 * pSB1AT3-J45120 pTet.BSMT
 * pSB1AK3-J45180 osmY.QPI.BSMT
 * have uncut and XP cut (69 ng/&mu;L)
 * pSB1AC3-J45700 (45120+45320) pTet.BSMT.pLac.SAGD
 * have uncut DNA (260ng/&mu;L)

To assemble
 
 * pSB1AC3-J45320.J45180
 * pSB1AK3-J45250.J45400
 * pSB1AK3-J45250.J45700
 * pSB1AK3-J45700.J45400

Smell tests

 * Use 4.4&mu;L stock IA-OH per 10mL culture
 * Use 40&mu;L 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