Team:Leeds/Results

From 2013.igem.org

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[[File:Leeds_Controlgrowthcurve.png|right|500px|Control growth-curve for Competent Cells, click for full image|link=https://static.igem.org/mediawiki/2013/e/e2/Leeds_Growthcurvecontrol.png|frameless]]
[[File:Leeds_Controlgrowthcurve.png|right|500px|Control growth-curve for Competent Cells, click for full image|link=https://static.igem.org/mediawiki/2013/e/e2/Leeds_Growthcurvecontrol.png|frameless]]
In this experiment we used our chosen expression cells and monitored their growth on SB media both in the presence of chloramphenicol and without chloramphenicol. As the bacteria do not contain our plasmid that codes for chloramphenicol resistance, we didn't expect them to grow when it was present. The graph produced from the absorbance measurements taken over time, shows the growth of the bacteria. It has a clear lag and log phase when the bacteria are grown in SB media only. But as expected, the graph shows that the bacteria do not grow in the presence of chloramphenicol. This graph will be used as a control and other growth curves of bacteria containing our device will be compared to it in order to determine whether the genes we inserted cause an effect on cell growth.
In this experiment we used our chosen expression cells and monitored their growth on SB media both in the presence of chloramphenicol and without chloramphenicol. As the bacteria do not contain our plasmid that codes for chloramphenicol resistance, we didn't expect them to grow when it was present. The graph produced from the absorbance measurements taken over time, shows the growth of the bacteria. It has a clear lag and log phase when the bacteria are grown in SB media only. But as expected, the graph shows that the bacteria do not grow in the presence of chloramphenicol. This graph will be used as a control and other growth curves of bacteria containing our device will be compared to it in order to determine whether the genes we inserted cause an effect on cell growth.
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==Agarose Gels For Digestions==
 
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When a digestion of DNA is undertaken, it is important that an Agarose gel is run afterwards to ensure it was successful before moving onto the next step in the experimental procedure.
 
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===Digestion of plasmid containing Green Fluorescent Protein===
 
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Here we digested the part [http://parts.igem.org/Part:BBa_K081012:Design K081012] with restriction enzyme EcoR1 and ran a gel to make sure the fragment(s) we obtained were of the correct length.
 
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[[File:Leeds_Agarosegelforgfp.png|left|300px|Agarose gel of GFP Digestion with EcoR1, click for full image|link=https://static.igem.org/mediawiki/2013/5/53/Leeds_DNAladdergraph.png|frameless]] This gel shows the bands obtained by EcoR1 digestion of BBa-K081012. There are 100bp ladders and 1kbp ladders in the first two wells. This gel was used to work out the size of the fragments obtained by plotting the results on a calibration graph.
 
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===DNA Ladder Calibration Graph===
 
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[[File:Leeds_DNAladdergraph.png|right|500px|DNA Ladder Calibration Graph for GFP Digestion, click for full image|frameless]]
 
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From this graph we can work out that one fragment is around 2800bp long, while the other appears to be 4000bp long - far longer than the plasmid at 2858bp long. Through digestion, it has been linearised by a single EcoR1 digest. The fragment corresponds to what we expect so our digest was successful, despite not all of the plasmid being digested.
 
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==Characterisation of Device 1==
==Characterisation of Device 1==

Revision as of 13:57, 21 August 2013

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Back to iGEM Main Page
Experimental Results
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 Results from the lab so far. Please note, many of the images have been uploaded at high resolution, and are best viewed from their file page. Access this by clicking the image. We may later introduce a gallery of the images to maximise the screen usage, but this is dependent upon Coffee-Induced coding by the Code Monkey.

Contents

Bacterial growth curves

We are monitoring the growth of our bacteria containing our devices to see if the genes we inserted have any effect on bacterial growth.

Control Bacterial Growth Curve

Control growth-curve for Competent Cells, click for full image

In this experiment we used our chosen expression cells and monitored their growth on SB media both in the presence of chloramphenicol and without chloramphenicol. As the bacteria do not contain our plasmid that codes for chloramphenicol resistance, we didn't expect them to grow when it was present. The graph produced from the absorbance measurements taken over time, shows the growth of the bacteria. It has a clear lag and log phase when the bacteria are grown in SB media only. But as expected, the graph shows that the bacteria do not grow in the presence of chloramphenicol. This graph will be used as a control and other growth curves of bacteria containing our device will be compared to it in order to determine whether the genes we inserted cause an effect on cell growth.

Characterisation of Device 1


Growth Curve


Growth curve for Device 1


Restriction Digests


File:Leeds BS1Restrictiondigests.png

Plate Imaging


Plate image for Device 1


Membrane Stress - Hydrophobic Stress


Membrane Stress - Changes in pH

A graph showing the effect pH has on the pCpxP Promoter


Membrane Stress- Atomic Force Microscopy


Characterisation of Device 2

Growth Curve

Growth curve for Device 2


Restriction Digests

Restriction digest for Device 2


Plate Imaging

Plate image for Device 2


Binding assay-washing beads


Binding assay-Confocal Microscopy


Characterisation of Device 3


Growth Curve

Growth curve for Device 3


Restriction Digests

Restriction Digests for Device 3


Plate Imaging

Plate image for Device 3


Binding Assay-Beads cause fluorescence?


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