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Team:Cambridge/Notebook/Week7
From 2009.igem.org
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(→MelA BioBrick) |
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====MelA BioBrick==== | ====MelA BioBrick==== | ||
| - | PCR done of the origonal MelA and the MelA | + | PCR done of the origonal MelA and the first MelA intermediate (without the second PstI site), to create the DNA for a PstI restriction digest. After PCR the DNA was run on a CYBRsafe-stained gel and then extracted with the QuiGen kit. A Restriction digest was carried out using the fastdigest buffer and PstI and left for three hours. |
We also tested the Primer C again, as it didn't work last time. Three PCR's were done: | We also tested the Primer C again, as it didn't work last time. Three PCR's were done: | ||
| - | *Primer F and Primer C used last time | + | *Primer F and Primer C (same aliquot as used last time, to check it wasn't just an experimental error on my part) |
*Primer F and a new aliquot of Primer C | *Primer F and a new aliquot of Primer C | ||
*Primer F and revortexed and re-aliquoted primer C from the origonal vial | *Primer F and revortexed and re-aliquoted primer C from the origonal vial | ||
| + | None of these produced any result - there is clearly an error in the primer that I ordered. | ||
===Dry Work=== | ===Dry Work=== | ||
Revision as of 06:00, 25 August 2009
Categories :
Project :
-
Overview
Sensitivity Tuner
--- Characterisation
--- Modelling
Colour Generators
--- Carotenoids (Orange/Red)
--- Melanin (Brown)
--- Violacein (Purple/Green)
The Future
Safety
Notebook :
Team Logistics :
Week 7
Monday
Wet Work
Amplification
Colony PCR of 8 colonies picked from 391 in pSB3K3 transformants, 8 colonies picked from 374 in pSB3K3 transformants, 8 colonies picked from 384 in pSB3K3, and 1 colony picked from 381 in pSB3K3 transformants confirmed to contain an insert of the right length. Overnight cultures of each of those colonies as well. No 374 constructs were found to contain inserts of the right length, however 4 384 constructs were of the correct length. The 381 construct was confirmed once again to be the correct length.
Overnight,
- Restirction digests of pSB3K3, 380, 382, 385, and 390.
- Colony PCR of 10 new 391 in pSB3K3 transformants, another 374 in pSB3K3 transformants
MelA BioBrick
PCR done of the origonal MelA and the first MelA intermediate (without the second PstI site), to create the DNA for a PstI restriction digest. After PCR the DNA was run on a CYBRsafe-stained gel and then extracted with the QuiGen kit. A Restriction digest was carried out using the fastdigest buffer and PstI and left for three hours.
We also tested the Primer C again, as it didn't work last time. Three PCR's were done:
- Primer F and Primer C (same aliquot as used last time, to check it wasn't just an experimental error on my part)
- Primer F and a new aliquot of Primer C
- Primer F and revortexed and re-aliquoted primer C from the origonal vial
None of these produced any result - there is clearly an error in the primer that I ordered.
Dry Work
We found the phzM and phzS pigments in the registry!
- BBa_I723024 (phzM)
- BBa_I723025 (phzS)
This means we can now start working on these pigments, connecting them to processing and logic systems.
