Team:Nevada/Notebook/Weeks1316

From 2011.igem.org

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(Week 13 - August 22nd-28th)
(Week 13 - August 22nd-28th)
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Enzymology<br>
Enzymology<br>
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This week we used the GC to analyze the fatty acids being produced by one of the BTE IQ cell samples (BTE13).  The fatty acids were extracted from cell supernatent using hexane and tested with a carbon disulfide solvent.  We used FAME standards ranging from C:8-C:19 as well as a C:19 internal standard for comparison.  Results proved positive as we were looking for C:12 FA's and there were C:8, C:10 and C:12 FA's present in the sample.  We suspect that the small chains could possibly be the larger C:12 chain being metabolized and we plan to further analyzed this data.  This was just a test or a pre-run and we did not use a negative control or test very many samples.  We plan to do a definite run next week on four new BTE samples as well as negative standards of wild-type IQ cells and a known concentration of a C:17 internal standard will be used for quantification.  Also this week we performed our third ethanol assay using the BioSystems Ethanol assay kit on 4 of the ADH/PDC IQ cell cultures.  We retrieved positive results for one of the samples (ACH/PDC17) with an ethanol production of 3.4mM.  We are planning to perform the ADH activity assay on this culture and four new samples next week.   
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This week we used the GC to analyze the fatty acids being produced by one of the BTE IQ cell samples (BTE13).  The fatty acids were extracted from cell supernatent using hexane and tested with a carbon disulfide solvent.  We used FAME standards ranging from C:8-C:19 as well as a C:19 internal standard for comparison.  Results proved positive as we were looking for C:12 FA's and there were C:8, C:10 and C:12 FA's present in the sample.  We suspect that the small chains could possibly be the larger C:12 chain being metabolized and we plan to further analyzed this data.  This was just a test or a pre-run and we did not use a negative control or test very many samples.  We plan to do a definite run next week on four new BTE samples as well as negative controls of wild-type IQ cells and a known concentration of a C:17 internal standard will be used for quantification.  Also this week we performed our third ethanol assay using the BioSystems Ethanol assay kit on 4 of the ADH/PDC IQ cell cultures.  We retrieved positive results for one of the samples (ACH/PDC17) with an ethanol production of 3.4mM.  We are planning to perform the ADH activity assay on this culture and four new samples next week.   
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Revision as of 16:19, 16 September 2011



Weeks 1-4    Weeks 5-8    Weeks 9-12    Weeks 13-16   

Calender Weeks 13-16
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Contents

Week 13 - August 22nd-28th

E. Coli

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Cyano

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Enzymology

This week we used the GC to analyze the fatty acids being produced by one of the BTE IQ cell samples (BTE13). The fatty acids were extracted from cell supernatent using hexane and tested with a carbon disulfide solvent. We used FAME standards ranging from C:8-C:19 as well as a C:19 internal standard for comparison. Results proved positive as we were looking for C:12 FA's and there were C:8, C:10 and C:12 FA's present in the sample. We suspect that the small chains could possibly be the larger C:12 chain being metabolized and we plan to further analyzed this data. This was just a test or a pre-run and we did not use a negative control or test very many samples. We plan to do a definite run next week on four new BTE samples as well as negative controls of wild-type IQ cells and a known concentration of a C:17 internal standard will be used for quantification. Also this week we performed our third ethanol assay using the BioSystems Ethanol assay kit on 4 of the ADH/PDC IQ cell cultures. We retrieved positive results for one of the samples (ACH/PDC17) with an ethanol production of 3.4mM. We are planning to perform the ADH activity assay on this culture and four new samples next week.



Media
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Week 14 - August 29th- September 4th

E. Coli

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Cyano

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Enzymology

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Media
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Week 15 - September 5th-11th

E. Coli
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Cyano

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Enzymology

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Media
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Week 16 - September 12th-18th

E. Coli
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Cyano

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Enzymology
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Media
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Week 17 - September 19th-25th

E. Coli
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Cyano

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Enzymology
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Media
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