console.js:26

Thursday June 30, 2016

Thursday, 6/30

Members Present:Andrea, Marc, Karim, Davesh, Kat, Hamed, Tam, Bohdan, Alex

LAB:

Morning:

- Miniprep and of 5mL overnight cultures of E.glowli and of L1:

	A	B	C	D
1	Sample name	260/280	260/230	concentration (ng/uL)
2	L1A	1.88	2.08	126.8
3	L1B	1.91	2.18	129.6
4	E. glowli A	1.88	1.51	48.6
5	E.glowli B	1.78	1.31	29.8

Table1

Miniprep results for L1 and for E.glowli. We have yet to run a get to try and confirm band sizes.

L1A - 30 nanodrop.jpg

L2A nanodrop - 30.jpg

E.glowli A nanodrop - 30.jpg

E.glowli B nanodrop - 30.jpg

- PCR amplification & nanodrop of the backbone from RFP:

	A	B	C	D
1	Sample name	260/280	260/230	concentration (ng/uL)
2	pSB1C3 backbone	1.45	0.59	712

Table2

Backbone PCR nanodrop.jpg

Afternoon:

●

Restriction enzyme digest (EcoRI, PstI) of LacZ PCR fragment with the samples from the 2-step PCR (29/06) and from the PCR at 68.1C (sample 5)

●

Restriction enzyme digest (EcoRI, PstI, DpnI) of the old backbone (24/06) and the new PCR backbone (30/06)

●

Ligation with the following, storing at -20 afterwards):

	A
1	Old bacbone - 2 step LacZ
2	Old backbone - sample 5 LacZ
3	New backbone - 2 step LacZ
4	New backbone - sample 5 LacZ

Table3

●

Grew E.glowli to OD600 ~0.2 and then induced with arabinose, and decreased the temperature to 30C to account for the temperature sensitivity of luciferase. BIOLUMINESCENCE OBSERVED.

E.glowli 1.jpg

E.glowli 2.jpeg

Administrative:

-Returned an order of ***insert herbicide name here*** that was mistakenly packaged with our L-Glutamic Acid Phosphate Salt, from Sigma Aldrich. Copy of the order transcript for the mistakenly packaged reagent can be found in the finance folder in WB403.

TO DO:

For the next week:

●

Test rubidium chloride competend cells CCD

●

Run gel on ligated (4x) and miniprepped parts.

●

Continue PCR amplifying IDT constructs.

LAB TEAM:

LAB MANAGERS:

Gmail correspondence:

Meetings/Notes:

References:

Team:Toronto/Notebook-w07-thu