Team:UC Davis/Notebook

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style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
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href="https://2009.igem.org/Team:UC_Davis/homepage2"><img alt=""
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href="https://2009.igem.org/Team:UC_Davis"><img alt=""
src="https://static.igem.org/mediawiki/2009/a/a4/UCDAVIS_PIC3.png"
src="https://static.igem.org/mediawiki/2009/a/a4/UCDAVIS_PIC3.png"
style="border: 0px solid ; width: 83px; height: 36px;"></a> </big></big></big></big></span></b><a
style="border: 0px solid ; width: 83px; height: 36px;"></a> </big></big></big></big></span></b><a
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href="https://2009.igem.org/Team:UC_Davis/About_Us1"><b
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style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
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href="https://2009.igem.org/Team:UC_Davis/Project1"><img alt=""
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href="https://2009.igem.org/Team:UC_Davis/Project"><img alt=""
src="https://static.igem.org/mediawiki/2009/b/b9/UCDAVIS_PIC8.png"
src="https://static.igem.org/mediawiki/2009/b/b9/UCDAVIS_PIC8.png"
style="border: 0px solid ; width: 78px; height: 36px;"></a> </big></big></big></big></span></b><b
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href="https://2009.igem.org/Team:UC_Davis/Notebook1"><img alt=""
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href="https://2009.igem.org/Team:UC_Davis/Notebook"><img alt=""
src="https://static.igem.org/mediawiki/2009/2/2f/UCDAVIS_PIC5.png"
src="https://static.igem.org/mediawiki/2009/2/2f/UCDAVIS_PIC5.png"
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style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
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href="https://2009.igem.org/Team:UC_Davis/Part1"><img alt=""
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src="https://static.igem.org/mediawiki/2009/a/a6/UCDAVIS_PIC6.png"
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style="border: 0px solid ; width: 78px; height: 37px;"></a>&nbsp;<a
style="border: 0px solid ; width: 78px; height: 37px;"></a>&nbsp;<a
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href="https://2009.igem.org/Team:UC_Davis/Contact1"><img alt=""
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href="https://2009.igem.org/Team:UC_Davis/Contacts_References"><img
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src="https://static.igem.org/mediawiki/2009/1/1d/UCDAVIS_PIC7.png"
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alt="" src="https://static.igem.org/mediawiki/2009/1/1d/UCDAVIS_PIC7.png"
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<hr style="width: 100%; height: 2px;"><b style="color: rgb(0, 0, 0);"><br>
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<hr style="width: 100%; height: 2px;"><b style="color: rgb(0, 0, 0);"><big><big><a
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</b></div>
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href="#july">July</a> &nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;
 +
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; <a href="#august">August</a>
 +
&nbsp;&nbsp; &nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp; &nbsp; <a
 +
href="#september">September</a>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
 +
<a href="#Results">Results</a><br>
 +
<br>
 +
<img style="width: 452px; height: 236px;" alt=""
 +
src="https://static.igem.org/mediawiki/2009/5/58/UCDAVIS_picture2.jpg"><br>
 +
</big></big></b>
 +
<hr style="width: 100%; height: 2px;">
 +
<p style="text-align: left;"><font face="Cambria" size="3"><big><big><span
 +
style="font-weight: bold;"><a name="july"></a>July:</span></big></big><br>
 +
&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;<span style="font-family: Calibri;">
 +
Before beginning the wet lab
 +
portion of our project, we have read numerous articles and journals
 +
relevant to our proposed project, and have utilized this information to
 +
guide our part designs.&nbsp; After significant research, design and
 +
redesign, we have settled on the final design for our two key parts,
 +
and plan for their construction.</span></font><span
 +
style="font-family: Calibri;">&nbsp;</span><br
 +
style="font-family: Calibri;">
 +
<font style="font-family: Calibri;" face="Cambria" size="3">&nbsp;&nbsp;&nbsp;
 +
&nbsp;&nbsp;&nbsp; We
 +
have begun ordering the necessary reagents for our work.&nbsp; This
 +
includes, selecting the parts required for our project from those
 +
provided by the iGEM parts distribution, oligonucleotides for
 +
extracting them, and for the synthesis of novel parts via Polymerase
 +
Chain Reaction, PCR, and the DNA synthesis of several novel parts from
 +
GENEART.</font><span style="font-family: Calibri;"> </span><font
 +
style="font-family: Calibri;" face="Cambria" size="3"><br>
 +
&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp; We have recently started the
 +
synthesis and assembly of the initial elements required for the
 +
secretion system.</font><span style="font-family: Calibri;">&nbsp;</span><br>
 +
</p>
 +
<hr style="width: 100%; height: 2px;">
 +
<div style="text-align: left;"><span style="font-family: Cambria;"></span><big
 +
style="font-weight: bold;"><big><a name="august"></a>August:</big></big><font
 +
face="Cambria" size="3"><br>
 +
&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp; <span
 +
style="font-family: Calibri;">After reading the numerous
 +
articles we dug up in regards to our project, we have designed our
 +
project; in blue print form of structural wet lab experimentation, as
 +
well as the techniques, materials, equipment, and time required to
 +
complete our project.</span></font><span style="font-family: Calibri;">
 +
</span><font style="font-family: Calibri;" face="Cambria" size="3"><br>
 +
&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp; The parts and a majority of the
 +
reagents we have ordered necessary for our work have arrived, and we
 +
are in the stages of conducting experiments to construct our
 +
parts.&nbsp; We have to construct, test, and prove through sequencing
 +
as well as other miscellaneous techniques that we truly have the parts
 +
we want in our constructs.&nbsp; So the progress for the month of
 +
August varies greatly on almost a daily basis.</font><span
 +
style="font-family: Calibri;">&nbsp;</span><br
 +
style="font-family: Calibri;">
 +
</div>
 +
<div style="text-align: left;"><font style="font-family: Calibri;"
 +
face="Cambria" size="3">&nbsp;&nbsp;&nbsp;
 +
&nbsp;&nbsp;&nbsp; Apart from the wet lab portion of our project, we
 +
have also begun the initial stages of building our wiki.&nbsp; We have
 +
begun to revise our information regarding the team, biomedical
 +
engineering, and how it is UC Davis’s inauguration year into the iGEM
 +
competition.</font><span style="font-family: Calibri;"> </span><br>
 +
<hr style="width: 100%; height: 2px;"><big><big><span
 +
style="font-weight: bold;"><a name="september"></a>September:<br>
 +
</span></big></big>
 +
<p><font face="Times New Roman" size="3">&nbsp;&nbsp;&nbsp; <span
 +
style="font-family: Calibri;">We are putting together the final pieces
 +
of our intended construct</span></font><font
 +
style="font-family: Calibri;" face="Times New Roman" size="3"> and
 +
have designed the tiny details of our experiments’ proper controls.The
 +
design itself is useless without making it as a physical part, so we
 +
are already building these simultaneously with our experimental
 +
constructs.</font><span style="font-family: Calibri;"> </span><br
 +
style="font-family: Calibri;">
 +
<span style="font-family: Calibri;">&nbsp;&nbsp;&nbsp; </span><font
 +
style="font-family: Calibri;" face="Times New Roman" size="3">The wiki
 +
is now up on the iGEM website and is being reviewed countless times to
 +
ensure accuracy and appeal.</font><font style="font-family: Calibri;"
 +
face="Times New Roman" size="3"><br>
 +
&nbsp;&nbsp;&nbsp; We have also designed team t-shirts, which are a
 +
tradition of iGEM, and are currently in the process of making them into
 +
print.</font><span style="font-family: Calibri;"></span><font
 +
style="font-family: Calibri;" face="Times New Roman" size="3">We are
 +
also making the presentation power point for presentation at the
 +
competition.</font><span style="font-family: Calibri;"> </span><font
 +
style="font-family: Calibri;" face="Times New Roman" size="3">We are
 +
also designed the poster for presentation at the competition.</font><span
 +
style="font-family: Calibri;"><br>
 +
</span></p>
 +
<hr style="width: 100%; height: 2px;">
 +
<div style="text-align: center;"><big><big><span
 +
style="font-weight: bold;"><a name="Results"></a>Results:<br>
 +
</span></big></big>
 +
<div style="text-align: left;"><big>Data collection strategy:<br>
 +
<br>
 +
Strain:<br>
 +
1. (no induction) ------&gt; Spin cells to seperate cells from media
 +
------&gt; Check for fluorescence or western blot with 6-His Tag<br>
 +
2. (induced with IPTG) ------&gt; </big><big>Spin cells to seperate
 +
cells from media ------&gt; </big><big>Check for fluorescence or</big><big>
 +
</big><big>western blot with 6-His Tag<br>
 +
<br>
 +
</big>
 +
<div style="text-align: center;"><big><img alt=""
 +
src="https://static.igem.org/mediawiki/2009/6/65/UCDAVIS_Exp_gel1.png"><br>
 +
</big>
 +
<div style="text-align: left;"><big><big><span
 +
style="font-weight: bold;"></span></big></big><br>
 +
</div>
 +
</div>
 +
<big><big><span style="font-weight: bold;"></span></big></big></div>
 +
<big><big><span style="font-weight: bold;"><br>
 +
</span></big></big></div>
 +
<span style="font-family: Calibri;">
 +
Whole cells were lyced and ran on this gel. The difference in protein
 +
concentrations are shown between the different plasmid containing
 +
cells. Each well was loaded with a cell containing the vectors labeled
 +
on top of the well.<br>
 +
</span>
 +
<div style="text-align: center;"><span style="font-family: Calibri;"><big><span
 +
style="font-weight: bold;"><br>
 +
INPNC_SS_GFP</span><br style="font-weight: bold;">
 +
<span style="font-weight: bold;">whole cell fluorescence spectrum</span></big><br>
 +
</span><span style="font-family: Calibri;"></span></div>
 +
<div style="text-align: center;"><span style="font-family: Calibri;"><img
 +
style="width: 764px; height: 485px;" alt=""
 +
src="https://static.igem.org/mediawiki/2009/7/73/UCDAVIS_Graph1.jpg"><br>
 +
Ex: 395 nm<br>
 +
Green = INPNC.SS.GFP induced<br>
 +
red = INPNC.SS.GFP uninduced<br>
 +
<br>
 +
</span><span style="font-family: Calibri;"></span></div>
 +
</div>
 +
</div>
</body>
</body>
</html>
</html>

Latest revision as of 01:19, 15 December 2009

12345final

 

July                August              September          Results



July:
        Before beginning the wet lab portion of our project, we have read numerous articles and journals relevant to our proposed project, and have utilized this information to guide our part designs.  After significant research, design and redesign, we have settled on the final design for our two key parts, and plan for their construction.
 
        We have begun ordering the necessary reagents for our work.  This includes, selecting the parts required for our project from those provided by the iGEM parts distribution, oligonucleotides for extracting them, and for the synthesis of novel parts via Polymerase Chain Reaction, PCR, and the DNA synthesis of several novel parts from GENEART.
        We have recently started the synthesis and assembly of the initial elements required for the secretion system.
 


August:
        After reading the numerous articles we dug up in regards to our project, we have designed our project; in blue print form of structural wet lab experimentation, as well as the techniques, materials, equipment, and time required to complete our project.

        The parts and a majority of the reagents we have ordered necessary for our work have arrived, and we are in the stages of conducting experiments to construct our parts.  We have to construct, test, and prove through sequencing as well as other miscellaneous techniques that we truly have the parts we want in our constructs.  So the progress for the month of August varies greatly on almost a daily basis.
 
        Apart from the wet lab portion of our project, we have also begun the initial stages of building our wiki.  We have begun to revise our information regarding the team, biomedical engineering, and how it is UC Davis’s inauguration year into the iGEM competition.

September:

    We are putting together the final pieces of our intended construct and have designed the tiny details of our experiments’ proper controls.The design itself is useless without making it as a physical part, so we are already building these simultaneously with our experimental constructs.
    The wiki is now up on the iGEM website and is being reviewed countless times to ensure accuracy and appeal.
    We have also designed team t-shirts, which are a tradition of iGEM, and are currently in the process of making them into print.
We are also making the presentation power point for presentation at the competition. We are also designed the poster for presentation at the competition.


Results:
Data collection strategy:

Strain:
1. (no induction) ------> Spin cells to seperate cells from media ------> Check for fluorescence or western blot with 6-His Tag
2. (induced with IPTG) ------>
Spin cells to seperate cells from media ------> Check for fluorescence or western blot with 6-His Tag




Whole cells were lyced and ran on this gel. The difference in protein concentrations are shown between the different plasmid containing cells. Each well was loaded with a cell containing the vectors labeled on top of the well.

INPNC_SS_GFP

whole cell fluorescence spectrum


Ex: 395 nm
Green = INPNC.SS.GFP induced
red = INPNC.SS.GFP uninduced