Difference between revisions of "Team:Toronto/Notebook-w07-thu"
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<li><a href="https://2016.igem.org/Team:Toronto/Integrated_Practices"><span>integrated_practices</span></a></li> | <li><a href="https://2016.igem.org/Team:Toronto/Integrated_Practices"><span>integrated_practices</span></a></li> | ||
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<li><a href="https://2016.igem.org/Team:Toronto/Model"><span>model</span></a></li> | <li><a href="https://2016.igem.org/Team:Toronto/Model"><span>model</span></a></li> | ||
Latest revision as of 06:56, 19 October 2016
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.
- 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
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.
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:
