Construction

From 2006.igem.org

Revision as of 03:44, 21 September 2006

September 20, 2006

Natalie, Andy:

• Prepared o/n growth of UT3 BCDE for mini-prep, DH5a for competent cells, and UT2 – 3,4,5, UT3 – 7,8,9 and DH5a x 2 (control) for making fluorescence measurements.
• Checked lengths of miniprep UT2 ABCDEF, UT3 A (possibly unreliable due to 2 day o/n growth), P0352 (2005), and P0152 (2005).

To-Do List:

• Make 24 competent cells
• Miniprep UT3 BCDE and test lengths
• Make more plates and re-plate P0452 (2005) – AK, and P0456 (2005) – AC
• Do measurements on UT2 and UT3
• Post up new protocols, include part length and resistance info in spreadsheet

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September 18, 2006

Charles:

• Only mini-prepped UT3 A because they were grown for two o/n (Just want to see what happens)
• Prepared new o/n for UT3 BCDE as well as 1 DH5a-z1, 2 UT2 and 2 UT3 for testing tomorrow. And 1 DH5a-z1 for making competent cells

To-Do List:

• Make 24 competent cells
• Test the lengths of UT2 ABCEDF, P0352 (2005), P0152 (2005), and UT3 BCDE
• Make more plates and re-plate P0452 (2005) – AK, and P0456 (2005) – AC
• Prepare new o/n for UT3 BCDE – for mini-prep
• Prepare o/n in the following quantities: 3 DH5a-z1, 3 UT2 and 3 UT3

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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 Registry
• 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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