Team:Leeds/Results

From 2013.igem.org

(Difference between revisions)
m
m
Line 12: Line 12:
We are monitoring the growth of our bacterial devices to see if any of the genes we have inserted have an effect on bacterial growth.
We are monitoring the growth of our bacterial devices to see if any of the genes we have inserted have an effect on bacterial growth.
===Control Bacterial Growth Curve===
===Control Bacterial Growth Curve===
-
[[File:Leeds_Growthcurvecontrol.png|right|450px|Control growth-curve for Competent Cells|frameless]]
+
[[File:Leeds_Growthcurvecontrol.png|right|500px|Control growth-curve for Competent Cells, click for full image|frameless]]
-
<br>This growth curve shows the lag phase and log phase of untransformed bacteria. The graph also shows that the bacteria do not grow in the presence of chloramphenecol. Our bacterial device will grow in the presence of chloramphenecol. We will use this graph as a comparison with our bacterial device.
+
This growth curve shows the lag phase and log phase of untransformed bacteria. The graph also shows that the bacteria do not grow in the presence of chloramphenecol. Our bacterial device will grow in the presence of chloramphenecol. We will use this graph as a comparison with our bacterial device.
<br style="clear:both" />
<br style="clear:both" />
==Agarose Gels For Digestions==
==Agarose Gels For Digestions==
Line 19: Line 19:
===Digestion of plasmid containing Green Flourescent Protein===
===Digestion of plasmid containing Green Flourescent Protein===
The part [http://parts.igem.org/Part:BBa_K081012:Design K081012] was digested with EcoR1 and a gel run to make sure the fragment(s)are of the correct length.
The part [http://parts.igem.org/Part:BBa_K081012:Design K081012] was digested with EcoR1 and a gel run to make sure the fragment(s)are of the correct length.
-
[[File:Leeds_Agarosegelforgfp.png|left|300px|Agarose gel of GFP Digestion with EcoR1|frameless]] This gel shows the bands obtained by EcoR1 digestion of BBa-K081012. There are 100bp ladders and 1kbp ladders. This gel was used to work out the size of the fragments obtained.
+
[[File:Leeds_Agarosegelforgfp.png|left|300px|Agarose gel of GFP Digestion with EcoR1, click for full image|frameless]] This gel shows the bands obtained by EcoR1 digestion of BBa-K081012. There are 100bp ladders and 1kbp ladders. This gel was used to work out the size of the fragments obtained.
-
<br>
+
<br style="clear:both" />
===DNA Ladder Calibration Graph===
===DNA Ladder Calibration Graph===
-
[[File:Leeds_DNAladdergraph.png|right|300px|DNA Ladder Calibration Graph for GFP Digestion|frameless]]From this graph we can work out that one fragment is around 3000bp long the other appears to be longer than the plasmid so we can assume this is undigested plasmid. The plasmid is 2858bp long. Through digestion it has been linearised by a single Ecor1 digest. The fragment corresponds to what we expect so our digest was succesful. Although not all of the plasmid was digested.
+
[[File:Leeds_DNAladdergraph.png|right|500px|DNA Ladder Calibration Graph for GFP Digestion, click for full image|frameless]]
-
<br>
+
From this graph we can work out that one fragment is around 3000bp long the other appears to be longer than the plasmid so we can assume this is undigested plasmid. The plasmid is 2858bp long. Through digestion it has been linearised by a single Ecor1 digest. The fragment corresponds to what we expect so our digest was succesful. Although not all of the plasmid was digested.
 +
<br style="clear:both" />
==Characterisation of Device 1==
==Characterisation of Device 1==
<br>
<br>
Line 29: Line 30:
<br>
<br>
[[File:Leeds_BS1growthcurve.png|left|300px|Growth curve for Device 1|frameless]]
[[File:Leeds_BS1growthcurve.png|left|300px|Growth curve for Device 1|frameless]]
-
<br>
+
<br style="clear:both" />
===Restriction Digests===
===Restriction Digests===
<br>
<br>
[[File:Leeds_BS1Restrictiondigests.png|left|300px|Restriction digest for Device 1|frameless]]
[[File:Leeds_BS1Restrictiondigests.png|left|300px|Restriction digest for Device 1|frameless]]
-
<br>
+
<br style="clear:both" />
===Plate Imaging===
===Plate Imaging===
<br>
<br>
[[File:Leeds_BS1plateimage.png|left|300px|Plate image for Device 1|frameless]]
[[File:Leeds_BS1plateimage.png|left|300px|Plate image for Device 1|frameless]]
-
<br>
+
<br style="clear:both" />
-
===Membrane Stress- Hydrophobic Stress===
+
===Membrane Stress - Hydrophobic Stress===
-
<br>
+
<br style="clear:both" />
-
===Membrane Stress- Changes in pH===
+
===Membrane Stress - Changes in pH===
-
<br>
+
[[File:Leeds_BS1pheffects.png|left|300px|A graph showing the effect pH has on the pCpxP Promoter|frameless]]
[[File:Leeds_BS1pheffects.png|left|300px|A graph showing the effect pH has on the pCpxP Promoter|frameless]]
-
<br>
+
<br style="clear:both" />
===Membrane Stress- Atomic Force Microscopy===
===Membrane Stress- Atomic Force Microscopy===
-
<br>
+
<br style="clear:both" />
==Characterisation of Device 2==
==Characterisation of Device 2==
-
<br>
 
===Growth Curve===
===Growth Curve===
-
<br>
 
[[File:Leeds_BS2Growthcurve.png|left|300px|Growth curve for Device 2|frameless]]
[[File:Leeds_BS2Growthcurve.png|left|300px|Growth curve for Device 2|frameless]]
-
<br>
+
<br style="clear:both" />
===Restriction Digests===
===Restriction Digests===
-
<br>
 
[[File:Leeds_BS2restrictiondigest.png|left|300px|Restriction digest for Device 2|frameless]]
[[File:Leeds_BS2restrictiondigest.png|left|300px|Restriction digest for Device 2|frameless]]
-
<br>
+
<br style="clear:both" />
===Plate Imaging===
===Plate Imaging===
-
<br>
 
[[File:Leeds_BS2plateimage.png|left|300px|Plate image for Device 2|frameless]]
[[File:Leeds_BS2plateimage.png|left|300px|Plate image for Device 2|frameless]]
-
<br>
+
<br style="clear:both" />
===Binding assay-washing beads===
===Binding assay-washing beads===
-
<br>
+
<br style="clear:both" />
===Binding assay-Confocal Microscopy===
===Binding assay-Confocal Microscopy===
-
<br>
+
<br style="clear:both" />
==Characterisation of Device 3==
==Characterisation of Device 3==
-
<br>
+
<br style="clear:both" />
===Growth Curve===
===Growth Curve===
-
<br>
 
[[File:Leeds_BS3Growthcurve.png|left|300px|Growth curve for Device 3|frameless]]
[[File:Leeds_BS3Growthcurve.png|left|300px|Growth curve for Device 3|frameless]]
-
<br>
+
<br style="clear:both" />
===Restriction Digests===
===Restriction Digests===
-
<br>
 
[[File:Leeds_BS3restrictiondigests.png|left|300px|Restriction Digests for Device 3|frameless]]
[[File:Leeds_BS3restrictiondigests.png|left|300px|Restriction Digests for Device 3|frameless]]
-
<br>
+
<br style="clear:both" />
===Plate Imaging===
===Plate Imaging===
-
<br>
 
[[File:Leeds_BS3plateimage.png|left|300px|Plate image for Device 3|frameless]]
[[File:Leeds_BS3plateimage.png|left|300px|Plate image for Device 3|frameless]]
-
<br>
+
<br style="clear:both" />
===Binding Assay-Beads cause fluorescence?===
===Binding Assay-Beads cause fluorescence?===
-
 
+
<br style="clear:both" />
}}
}}

Revision as of 18:57, 13 August 2013

Parky roof.png
Parky top.png
Back to iGEM Main Page
Experimental Results
Parky main.png
iGEM Leeds Facebook
iGEM Leeds Twitter
iGEM Leeds Youtube
Leeds socialbanneredge.png
Parky tower.png
 Here are Our Results from the lab so far

Bacterial growth curves

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

Control Bacterial Growth Curve

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

This growth curve shows the lag phase and log phase of untransformed bacteria. The graph also shows that the bacteria do not grow in the presence of chloramphenecol. Our bacterial device will grow in the presence of chloramphenecol. We will use this graph as a comparison with our bacterial device.

Agarose Gels For Digestions

For every Digestion we do we will run a gel to ensure the digestion was successful, and use gel extraction kits to obtain purified DNA.

Digestion of plasmid containing Green Flourescent Protein

The part [http://parts.igem.org/Part:BBa_K081012:Design K081012] was digested with EcoR1 and a gel run to make sure the fragment(s)are of the correct length.

Agarose gel of GFP Digestion with EcoR1, click for full image
This gel shows the bands obtained by EcoR1 digestion of BBa-K081012. There are 100bp ladders and 1kbp ladders. This gel was used to work out the size of the fragments obtained.


DNA Ladder Calibration Graph

DNA Ladder Calibration Graph for GFP Digestion, click for full image

From this graph we can work out that one fragment is around 3000bp long the other appears to be longer than the plasmid so we can assume this is undigested plasmid. The plasmid is 2858bp long. Through digestion it has been linearised by a single Ecor1 digest. The fragment corresponds to what we expect so our digest was succesful. Although not all of the plasmid was digested.

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?


Parky base.png
Geneious, our fine sponsors and suppliers of software Bioline, our fine sponsors and suppliers of equipment Qiagen, our fine sponsors and suppliers of PCR kits
Bangs Laboratories, our fine sponsors and suppliers of silica beads
Leeds Homepage
Retrieved from "http://2013.igem.org/Team:Leeds/Results"