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 transformed with the ''glgC16'' BioBrick but grown in a glucose-free medium. These should produce a higher level of glycogen than the control.
+
* '''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 transformed with the ''glgC16'' Biobrick and grown in a high-glucose medium. These should produce the highest level of glycogen.
+
* '''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

Edinburgh=Glycogen-Assay3.jpg

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.