Team:Freiburg/Description

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(Difference between revisions)
(The temperature sensitive lysis cassette)
(The temperature sensitive lysis cassette)
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What we managed to do, was (finally!) clone it some days before the freeze and "blindly" make some preliminary OD measurements days in advance of sequencing in order to detect lysis [and, of course, be able to provide some data] since the sequencing company could not deliver results until some 18 hours before the Wiki freeze...<br>
What we managed to do, was (finally!) clone it some days before the freeze and "blindly" make some preliminary OD measurements days in advance of sequencing in order to detect lysis [and, of course, be able to provide some data] since the sequencing company could not deliver results until some 18 hours before the Wiki freeze...<br>
What these results showed, was indeed lysis but the correlation with temperature control was not what we expected it to be.
What these results showed, was indeed lysis but the correlation with temperature control was not what we expected it to be.
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Here is what we managed to measure [click for a bigger Image] (again, this was a blind preliminary measurement days in advance of sequencing without any real planning)<br>
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Here is what we managed to measure [click for a bigger Image] (again, this was a blind preliminary measurement days in advance of sequencing without any real planning)
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<br>
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[[File:Freiburg11 LysisCassetteODmeasurement.jpg|800px]]
[[File:Freiburg11 LysisCassetteODmeasurement.jpg|800px]]
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Nr1 proved to be only the temperature sensitive promoter (BBa_K608351)<br>
Nr1 proved to be only the temperature sensitive promoter (BBa_K608351)<br>
Nr2, on the other side, proved to be only the lysis cassette (BBa_K608352), which makes for a cool set of controls just by chance :)<br>
Nr2, on the other side, proved to be only the lysis cassette (BBa_K608352), which makes for a cool set of controls just by chance :)<br>
Nr3 is indeed the composite part of BBa_K608351 + BBa_K608352.<br>
Nr3 is indeed the composite part of BBa_K608351 + BBa_K608352.<br>
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Since we cannot upload ab1 files, here are the corresponding .gb files (right click to download, please bear in mind that since they are direct basecalls of the ab1 files, no pruning has been done. Also there are some bps missing in the middle for GATC Biotech GmbH could obviously not manage to sequence 2200bp with fw and rev primers...)<br>  
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:Since we cannot upload ab1 files, here are the corresponding .gb files (right click to download, please bear in mind that since they are direct basecalls of the ab1 files, no pruning has been done. Also there are some bps missing in the middle for GATC Biotech GmbH could obviously not manage to sequence 2200bp with fw and rev primers...)<br>  
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[[File:Freiburg11 S48+S66 Nr3-P8,fw.ab1 basecalls .gb]]<br>
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::[[File:Freiburg11 S48+S66 Nr3-P8,fw.ab1 basecalls .gb]]<br>
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[[File:Freiburg11 S48+S66 Nr3-P9,rev.ab1 basecalls .gb]]<br>
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::[[File:Freiburg11 S48+S66 Nr3-P9,rev.ab1 basecalls .gb]]<br>
The measurement of the optical density shows that the lysis cassette is indeed controlled by the promoter, but a leaky expression is obvious and further statements cannot be made since we only grew the cells at 30°C and not 28°C or even 25°C to tighten the putatively leaky expression of the cassette
The measurement of the optical density shows that the lysis cassette is indeed controlled by the promoter, but a leaky expression is obvious and further statements cannot be made since we only grew the cells at 30°C and not 28°C or even 25°C to tighten the putatively leaky expression of the cassette

Revision as of 13:37, 21 September 2011


This is the wiki page
of the Freiburger student
team competing for iGEM 2011.
Thank you for your interest!