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Team:Nevada/Notebook/Weeks912
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Enzymology<br> | Enzymology<br> | ||
| - | + | This week we performed our time course growth on the positive culture sample taking samples every two hours with an overnight culture as well. We performed the FFA assay on every time course sample using the BioAssay Systems FFA assay kit with slight modifications to the manufactures protocol. The results were negative for all samples which could have possibly been due to improper storage of the assay kit. Another assay kit was ordered as samples were to be further tested. <br> | |
</font> <br> | </font> <br> | ||
Media<br> | Media<br> | ||
Revision as of 20:59, 27 August 2011
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Calender Weeks 9-12
To edit on a specific week. Click on the edit button corresponding to the week.
Contents |
Week 9 - July 25th-31st
E. Coli
Comment Here
Cyano
7/25/11: Chloramphenicol vector was digested with EcoRI and checked on 0.7% gel. The results for this gel were good. ThiE/Topo Vector was digest with EcoRI and PstI and checked on a 0.7% gel. The gel was not very clean, so we will re-run tomorrow (7/26).
The PetBD and GLF PCR was done at 63.5C. Positive control/primer controls were added to the PCR reaction.
After confirmation of ThiE it will be put into plasmid PSB1C3 and sequenced.
7/26/11: Gels for PCR products of PetBD and GLF were run with no results. We believe this is due to errors in the master mix.
ThiE digest was re-run on a 0.7% gel. The gel showed that the ThiE was not digested. (7/27).
Ran PCR reaction on ThiE gel purification 7/18/11 with positive control and primer control at 54.3C for Topo Cloning tomorrow (7/27).
7/27/11: Ran gel on + control, Primer control and ThiE to be used for Topo cloning. Gel was completely empty aside from ladders. Did digest on ThiE in Topo Vector with ecoRI. Results showed that we have an empty topo Vector on all three digests (#'s 2, 3 and 5).
Enzymology
This week we performed our time course growth on the positive culture sample taking samples every two hours with an overnight culture as well. We performed the FFA assay on every time course sample using the BioAssay Systems FFA assay kit with slight modifications to the manufactures protocol. The results were negative for all samples which could have possibly been due to improper storage of the assay kit. Another assay kit was ordered as samples were to be further tested.
Media
Comment Here
Week 10 - August 1st-7th
E. Coli
Pyruvate Decarboxylase & Alcohol Dehydrogenase
PDC/ADH was re-digested with XbaI/PstI and run on a 1% agarose gel to confirm digestion. Bands obtained confirmed PDC/ADH (3054bp), and pSB1C3 (2070bp). PDC/ADH-XbaI/PstI and σ70/pSB1A3-SpeI/PstI were re-ligated and transformed into NEB10 β cells.
Cyano
Comment Here
Enzymology
Comment Here
Media
Comment Here
Week 11 - August 8th-14th
E. Coli
Pyruvate Decarboxylase & Alcohol Dehydrogenase
Single colonies from LB-Amp plates were selected and tested on LB-Amp and LB-Chlor plates. Liquid cultures grown in LB-Amp and minipreps and nanodrop analysis performed.
To verify presence of PDC/ADH genes with the σ70 constitutive promoter, 0.5ug of PDC/ADH/σ70/pSB1A3 was digested with EcoRI and PstI. Digests were run on a 1% agarose gel, and bands confirmed σ70/PDC/ADH (3089bp) and pSB1A3 (2155bp).
Cyano
Comment Here
Enzymology
Comment Here
Media
Comment Here
Week 12 - August 15th-21st
E. Coli
Comment Here
Cyano
Comment Here
Enzymology
Comment Here
Media
Comment Here
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/* Wiki Hacks - START */
/* Author: Pieter van Boheemen */
/* Team: TU Delft */
/* Thanks guys - Bill Collins */
/* +1 - Douglas Watson */
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