Construction

From 2006.igem.org

September 16, 2006

Charles:

• Made 4 replications of UT2 in 200 uM, 100 uM, 50 uM, 25 uM and 0 uM of IPTG.
  
  
  
  • OD of cells is too high, need to be <= 1.2
  • Need a control of untransformed DH5a-z1 to compare the relative fluorescence
• Made frozen stock (2 vials each): UT01, I13507 (2005), B0015 (2005), B0031 (2005)
• Mini-prepped UT2 ABCEDF, P0352 (2005) and P0152 (2005)

To-Do List:

• Test the lengths of UT2 ABCEDF, P0352 (2005) and P0152 (2005)
  • Prepare an o/n growth of the “best” UT2 for testing
  • Prepare an o/n growth of regular DH5a-z1
• Mini-prep UT03 ABCD and test their lengths along with P00352 (2005) and P0152 (2005)
• Re-plate P0452 (2005) - AK, and P0456 (2005) - AC
• Make 24 competent cells

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September 15, 2006

Andy, Charles, Natalie, Stan, Elliott:

• Checked and transformed I+J+I plasmids (Bands: ~6000-5000, 3500-2500 (smear))
• Made competent cells (although, may have to repeat)
• Prepared o/n growths of I+J+E (12 vials), P0152 (2005) and P0352 (2005). P0456 (2005) and P0452 (2005) did not seem to grow very well so left them in the incubator for a 2nd night
• Made 8 Amp plates

To-Do List:

• Mini-prep P0152 (2005), P0352 (2005) and 6 of the I+J+E
• Try to get J04450 (2006) working again.
• Check lengths of R0011 (2005/6), if no good, then obtain R0010 (2005/6) from Regsitry
• Make freezer stock (2 each) of I+J, B0015 (2005), B0031 (2005), and I13507 (2005) and put in -80C freezer stock
• Test the fluorescence of I+J+E

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