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Team:Edinburgh/Results/Glycogen3
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This quantitative assay compared the amount of glycogen produced by: | This quantitative assay compared the amount of glycogen produced by: | ||
* '''Sample 1 (Control):''' ''E. coli'' cells with no modification to the glycogen production system and grown in a medium which does not promote glycogen formation. These should have a low, basal level of glycogen. | * '''Sample 1 (Control):''' ''E. coli'' cells with no modification to the glycogen production system and grown in a medium which does not promote glycogen formation. These should have a low, basal level of glycogen. | ||
| - | * '''Sample 2:''' ''E. coli'' cells | + | * '''Sample 2:''' ''E. coli'' cells overexpressing ''glgC16'' BioBrick and grown in a glucose-free medium. These should produce a higher level of glycogen than the control. |
| - | * '''Sample 3:''' ''E. coli'' cells | + | * '''Sample 3:''' ''E. coli'' cells overexpressing with the ''glgC16'' Biobrick and grown in a high-glucose medium. These should produce the highest level of glycogen. |
Results from [[Team:Edinburgh/Results/Glycogen2|Glycogen Assay 2 (Qualitative)]] were in line with these expectations. | Results from [[Team:Edinburgh/Results/Glycogen2|Glycogen Assay 2 (Qualitative)]] were in line with these expectations. | ||
Revision as of 22:42, 27 October 2008
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Glycogen Assay 3 (Quantitative: Raman)
Raman spectroscopy was performed by Dr Rabah Mouras.
Experiment Design
This quantitative assay compared the amount of glycogen produced by:
- Sample 1 (Control): E. coli cells with no modification to the glycogen production system and grown in a medium which does not promote glycogen formation. These should have a low, basal level of glycogen.
- Sample 2: E. coli cells overexpressing glgC16 BioBrick and grown in a glucose-free medium. These should produce a higher level of glycogen than the control.
- Sample 3: E. coli cells overexpressing with the glgC16 Biobrick and grown in a high-glucose medium. These should produce the highest level of glycogen.
Results from Glycogen Assay 2 (Qualitative) were in line with these expectations.
The three samples were analysed by Raman spectroscopy to determine the glycogen:protein ratio. Glycogen results in a peak at a Raman shift of 478cm-1, so the Raman intensity of this peak was measured for each sample and compared to the Phenylalanine peak at 1003cm-1.
Results
The results confirm our expectations. Control cells (S1) produced the least amount of glycogen and glgC16 cells grown in high-glucose medium (S3) the most. From the intensity of the glycogen peak, the concentration of glycogen in S3 is estimated to be 3~4 times more than in S2.
We may thus conclude that the glgC16 BioBrick results in significantly increased production of glycogen, especially in a high-glucose medium.
