Team:Toronto/Notebook-w10-fri
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Friday - July 22
Friday, 7/22
Members Present: Kat, Tam, Hamed, Bohdan
LAB:
Morning:
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Made 2 x 250ml cultures of BL21 in 2x YPTG. Put in the orbital shaker at 300rpm
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Measuring the OD after every half hour
A | B | |
1 | Time point | OD600 (A) |
2 | 1 | 0.027 |
3 | 2 | 0.041 |
4 | 3 | 0.101 |
5 | 4 | 0.274 |
6 | 5 | |
7 | 6 | |
8 | 7 | |
9 | 8 | |
Table1
Afternoon:
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Completed first day of S30 protocol (centrifugation, 3xwashes with buffer A, flash freezing with liquid nitrogen); there are now four falcon tubes with washed cells in the -80 freezer.
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Carried out gel extraction protocol on backbone (we made sure anything above 2.5kb [i.e. RFP] was disposed of).
A | B | C | D | |
1 | SAMPLE | 260/280 | 260/230 | ng/uL |
2 | Backbone 1 | 2.49 | 0.02 | 12 |
3 | Backbone 2 | 2.05 | 0.3 | 17.1 |
4 | Backbone 3 | 2.56 | 0.03 | 19.6 |
5 | Backbone 4 | 4.49 | 0.05 | 21.8 |
Table4
Note that the backbone samples were labelled from the four slices of gel extracted, going from left to right. Therefore Backbone 1 and backbone 4 refer to the leftmost and rightmost slices, respectively.
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Transformed CCD cells using constructs ligated overnight (transformed with 2ul and 5ul samples of the ligated constructs; CjBlue was used as control to avoid red colonies):
A | B | |
1 | 2ul | 5ul |
2 | RFP+ | RFP+ |
3 | RFP- | RFP- |
4 | Kit+ | Kit+ |
5 | Kit- | Kit- |
6 | CjBlue (Contro) | |
Table3
(RFP: BB from PCR amplification of RFP; Kit: BB from PCR amplification of Kit BB; +: used rSAP on BB; -: did not use rSAP on BB)
Administrative:
Items purchased:
A | B | C | D | E | F | |
1 | Name | SKU | Unit Price | Quantity | Total (incl. tax) | Status |
2 | Liquid Nitrogen (1L) | 2384 | $1.71 | 1 | 1.91 | temporarily available |
Table2
TO DO:
For the next day:
LAB TEAM:
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Pray to the lab gods that our ligations worked
