IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-8-6

To-do

1. Make frozen stocks of the KaiA+bkb and KaiB+bkb transformants that we grew up in liquid culture yesterday
2. Miniprep the above
3. (Maybe) Do an X-P digest of the KaiA+bkb and KaiB+bkb plasmids (though we may want to wait and do this along with the other KaiA+bkb and KaiB+bkb ligation transformants)
4. Inoculate the KaiA+bkb and KaiB+bkb transformants from yesterday's ligation
5. Add CIP to the E-S digest of LC vector
6. Run the digests from yesterday on a gel (E-S digest of LC vector, X-P digest of J04450+bkb)
7. Gel-extract and purify the LC vector and J04450 insert if they exist
8. (maybe) Perform a ligation and transformation of the J04450 insert into LC vector
9. Do 12 minipreps for Mingming
10. Check sequencing on the Genewiz website (the orders have not been processed)

KaiA/B + bkb inoculation

We found 10 colonies on the KaiA+bkb plate and 5 colonies on the KaiB+bkb plate. We inoculated all 15 colonies in liquid culture (8 mL LB, 8 uL Amp) and placed them in the 37C shaker around 1830.

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