Construction

Revision as of 20:42, 18 September 2006 by Chuck (Talk | contribs)

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
Repeat previous experiment using varying concentration of IPTG for both UT2 and UT3 with a DH5a-z1 control

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Charles:

Made 4 replications of UT2 in 200 uM, 100 uM, 50 uM, 25 uM and 0 uM of IPTG.

Module01_Test01_Graph.jpg
- 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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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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