Team:Calgary/6 July 2010


Revision as of 20:43, 6 July 2010 by Cktang (Talk | contribs)

Tuesday July 6, 2010

Himika:* Today I reran the transformation that I did yesterday July 5th where I constructed E0040 and pSB1AC3 and plate it on Chloromphenicol.

  • I also miniprepped the overnight cultures from last night. I10504 and I10507. The only cultures that grew were I10504 and I miniprepped 6 colonies from 1 plate.
  • I also constructed E1010 with B0015.
  • Procedure:
    • Insert
    • E1010 (pSB2k3) cut with EcoI/SpeI 5ul DNA
    • Buffer I was used 3.5ul
    • 0.5 ul of EcoRI and 0.5 ul of SpeI
    • 25.5 uL of water
    • Vector
    • B0015 (pSB1AK3) cut with EcoI/XbaI 5ul DNA
    • Buffer I was used 3.5ul
    • 0.5 ul of EcoRI and 0.5 ul of XbaI
    • 25.5 uL of water
  • The vector was phosphatased with 5ul water, 4ul Buffer and 1ul phosphatase. The solution was left in 37 degree celcius for 30 minutes and heat killed for 10 minutes in 65 degree celcius.
  • 5ul of the vector and the insert in a tube and add 10 ul of quick ligase buffer and 1 ul of quick ligase. The solution was left in room temperature for 10 minutes and transformed.
  • I also did research on the ibpA/ ibpB pathways in order to present on to Dr. Schryvers on Friday.
  • I also overnight cultured I10507 and plated the contstruct.

No notebook page exists for this date. Sorry!