Difference between revisions of "Team:UPF CRG Barcelona/InterLab"

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{{UPF_CRG_Barcelona}}
 
{{UPF_CRG_Barcelona}}
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<HTML>
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<HEAD>
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<META HTTP-EQUIV="CONTENT-TYPE" CONTENT="text/html; charset=utf-8">
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<TITLE></TITLE>
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<STYLE TYPE="text/css">
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@page {
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margin-left: 1.18in;
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margin-right: 1.18in;
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margin-top: 0.98in;
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color: #0563c1;
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so-language: zxx
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}
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</HEAD>
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<BODY LINK="#0563c1" DIR="LTR">
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<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
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<FONT FACE="Helvetica, serif">
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<FONT SIZE=4 STYLE="font-size: 16pt">
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<SPAN LANG="en-US">
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<B>5</B>
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</SPAN>
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</FONT>
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</FONT>
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<SUP>
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<FONT FACE="Helvetica, serif">
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<FONT SIZE=4 STYLE="font-size: 16pt">
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<SPAN LANG="en-US">
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<B>th</B>
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</SPAN>
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</FONT>
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</FONT>
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</SUP>
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<FONT FACE="Helvetica, serif">
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<FONT SIZE=4 STYLE="font-size: 16pt">
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<SPAN LANG="en-US">
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<B>
 +
INTERLAB STUDY</B>
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</SPAN>
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</FONT>
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</FONT>
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</P>
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<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
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<FONT FACE="Helvetica, serif">
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<SPAN LANG="en-US">
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<B>Introduction</B>
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</SPAN>
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</FONT>
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</P>
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<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
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<FONT FACE="Helvetica, serif">
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<FONT SIZE=2 STYLE="font-size: 11pt">
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<SPAN LANG="en-US">The aim is to develop a reliable and repeatable measurement based on cell number, fluorescence, absorbance (optical density)
 +
and colony formation units (CFUs). iGEM teams collaborate in measuring these parameters following the same protocol
 +
to obtain a way to have accurate and reliable measurements, which are essential for all sciences, including synthetic
 +
biology. </SPAN>
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</FONT>
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</FONT>
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</P>
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<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
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<FONT FACE="Helvetica, serif">
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<FONT SIZE=2 STYLE="font-size: 11pt">
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<SPAN LANG="en-US">The center of Interlab has been always the green fluorescent protein (GFP), one of the biological markers most used in
 +
synthetic biology. </SPAN>
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</FONT>
 +
</FONT>
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</P>
 +
<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
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<FONT FACE="Helvetica, serif">
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<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<SPAN LANG="en-US">The goal of the fifth edition of Interlab is to discover the sources of variability in measurements and be able to correct
 +
them, so the measurements taken in different labs will not be variable anymore. </SPAN>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
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<SPAN LANG="en-US">This year question is: </SPAN>
 +
</FONT>
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</FONT>
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<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<SPAN LANG="en-US">
 +
<B>Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or CFUs instead
 +
of OD? </B>
 +
</SPAN>
 +
</FONT>
 +
</FONT>
 +
</P>
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<P LANG="en-US" ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<BR>
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</P>
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<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<FONT FACE="Helvetica, serif">
 +
<SPAN LANG="en-US">
 +
<B>Materials and Methods</B>
 +
</SPAN>
 +
</FONT>
 +
</P>
 +
<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<SPAN LANG="en-US">Before the starting the experimental part, a plate reader was needed. Due to the lack of plate reader in our laboratory,
 +
it was kindly asked to Proteomics and Protein Chemistry Unit (DCEXS-UPF, PRBB) to use their equipment. The plate reader
 +
is the Synergy HTX Multimode Reader from Biotek, it can measure both absorbance and fluorescence. It has pathlength
 +
correction, which was not disconnected. It has control over temperature and it was set as room temperature (around 24-25
 +
Celsius degrees). The excitation filter was 485/20 nm and the emission filter 528/20 nm and bottom optics were used.
 +
Moreover, the plates were black and flat-bottomed. </SPAN>
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</FONT>
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</FONT>
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</P>
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<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
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<A NAME="_GoBack"></A>
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<FONT FACE="Helvetica, serif">
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<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<SPAN LANG="en-US">Eight plasmids need to be characterized in DH5-alpha E.coli strain in order to follow the protocol. The strain was obtained
 +
by collaboration with BIO-IQS iGEM team. The plasmids are the following: BBa_R0040, BBa_I20270, BBa_J3604000, BBA_J364001,
 +
BBa_J364002, BBa_J364007, BBa_J364008, BBa_J364009. </SPAN>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<SPAN LANG="en-US">The materials used over the protocol are the same ones specified in the
 +
<A HREF="https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf">iGEM 2018 Interlab Study Protocol</A>.
 +
</SPAN>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<FONT FACE="Helvetica, serif">
 +
<SPAN LANG="en-US">
 +
<B>Results</B>
 +
</SPAN>
 +
</FONT>
 +
</P>
 +
<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<FONT FACE="Helvetica, serif">
 +
<SPAN LANG="en-US">
 +
<B>Calibration</B>
 +
</SPAN>
 +
</FONT>
 +
</P>
 +
<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<FONT FACE="Helvetica, serif">
 +
<SPAN LANG="en-US">
 +
<U>
 +
<B>OD600 Reference Point</B>
 +
</U>
 +
</SPAN>
 +
</FONT>
 +
</P>
 +
<CENTER>
 +
<TABLE style="width: 600px">
 +
<TR>
 +
<TD WIDTH=156 HEIGHT=3 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<BR>
 +
</P>
 +
</TD>
 +
<TD WIDTH=106 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>LUDOX CL-X
 +
</B>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=71 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>H</B>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
<FONT COLOR="#000000">
 +
<SUB>
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>2</B>
 +
</FONT>
 +
</FONT>
 +
</SUB>
 +
</FONT>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>O</B>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=156 HEIGHT=6 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>Replicate 1
 +
</B>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=106 BGCOLOR="#e8f3ff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2>0,056</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=71 BGCOLOR="#e8f3ff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2>0,034</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=156 HEIGHT=6 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>Replicate 2
 +
</B>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=106 BGCOLOR="#e8f3ff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2>0,056</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=71 BGCOLOR="#e8f3ff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2>0,034</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=156 HEIGHT=6 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>Replicate 3
 +
</B>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=106 BGCOLOR="#e8f3ff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2>0,056</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=71 BGCOLOR="#e8f3ff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2>0,034</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=156 HEIGHT=6 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>Replicate 4
 +
</B>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=106 BGCOLOR="#e8f3ff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2>0,056</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=71 BGCOLOR="#e8f3ff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2>0,034</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=156 HEIGHT=6 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>Arith. Mean
 +
</B>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=106 BGCOLOR="#127abd" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">0,056</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=71 BGCOLOR="#127abd" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">0,034</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=156 HEIGHT=6 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>Corrected Abs600
 +
</B>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=106 BGCOLOR="#127abd" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">0,022</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=71 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<BR>
 +
</P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=156 HEIGHT=6 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>Reference OD600
 +
</B>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=106 BGCOLOR="#127abd" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">0,063</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=71 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<BR>
 +
</P>
 +
</TD>
 +
</TR>
 +
<TR>
 +
<TD WIDTH=156 HEIGHT=6 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<B>OD600/Abs600</B>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=106 BGCOLOR="#51bdb2" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER>
 +
<FONT COLOR="#000000">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">2,864</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
</TD>
 +
<TD WIDTH=71 BGCOLOR="#ffffff" STYLE="border: none; padding: 0in">
 +
<P ALIGN=CENTER STYLE="page-break-after: avoid">
 +
<BR>
 +
</P>
 +
</TD>
 +
</TR>
 +
</TABLE>
 +
</CENTER>
 +
<P ALIGN=CENTER STYLE="margin-bottom: 0.14in">
 +
<FONT COLOR="#44546a">
 +
<FONT SIZE=2 STYLE="font-size: 9pt">
 +
<FONT COLOR="#000000">
 +
<SPAN LANG="en-US">
 +
<SPAN STYLE="font-style: normal">Table 1 | Date from the OD600 Reference Point in calibration 1.</SPAN>
 +
</SPAN>
 +
</FONT>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<FONT FACE="Helvetica, serif">
 +
<SPAN LANG="en-US">
 +
<U>
 +
<B>Particle Standard Curve</B>
 +
</U>
 +
</SPAN>
 +
</FONT>
 +
</P>
 +
<P ALIGN=CENTER STYLE="margin-bottom: 0in; line-height: 150%">
 +
<IMG src="https://static.igem.org/mediawiki/2018/8/86/T--UPF_CRG_Barcelona--interlabcalibration2.png" NAME="Picture 4" ALIGN=BOTTOM WIDTH=567 HEIGHT=556 BORDER=0>
 +
</P>
 +
<P LANG="en-US" ALIGN=CENTER STYLE="margin-bottom: 0in; line-height: 150%">
 +
<BR>
 +
</P>
 +
<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<FONT FACE="Helvetica, serif">
 +
<SPAN LANG="en-US">
 +
<U>
 +
<B>Fluorescence Standard Curve</B>
 +
</U>
 +
</SPAN>
 +
</FONT>
 +
</P>
 +
<P ALIGN=CENTER STYLE="margin-bottom: 0in; line-height: 150%">
 +
<IMG src="https://static.igem.org/mediawiki/2018/6/61/T--UPF_CRG_Barcelona--interlabcalibration3.png" NAME="Picture 5" ALIGN=BOTTOM WIDTH=567 HEIGHT=575 BORDER=0>
 +
</P>
 +
<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<SPAN LANG="en-US">This calibration needs to be repeated because of the low R</SPAN>
 +
</FONT>
 +
</FONT>
 +
<SUP>
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<SPAN LANG="en-US">2</SPAN>
 +
</FONT>
 +
</FONT>
 +
</SUP>
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<SPAN LANG="en-US">
 +
(R</SPAN>
 +
</FONT>
 +
</FONT>
 +
<SUP>
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<SPAN LANG="en-US">2</SPAN>
 +
</FONT>
 +
</FONT>
 +
</SUP>
 +
<FONT FACE="Helvetica, serif">
 +
<FONT SIZE=2 STYLE="font-size: 11pt">
 +
<SPAN LANG="en-US">=0,8547). But it can be caused by the plate reader since it was unable to read some of the biggest concentrations in
 +
the plate (10 uM of fluorescein). </SPAN>
 +
</FONT>
 +
</FONT>
 +
</P>
 +
<P ALIGN=JUSTIFY STYLE="margin-bottom: 0in; line-height: 150%">
 +
<FONT FACE="Helvetica, serif">
 +
<SPAN LANG="en-US">
 +
<B>Measurement</B>
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Revision as of 17:01, 24 July 2018

Wiki

5 th INTERLAB STUDY

Introduction

The aim is to develop a reliable and repeatable measurement based on cell number, fluorescence, absorbance (optical density) and colony formation units (CFUs). iGEM teams collaborate in measuring these parameters following the same protocol to obtain a way to have accurate and reliable measurements, which are essential for all sciences, including synthetic biology.

The center of Interlab has been always the green fluorescent protein (GFP), one of the biological markers most used in synthetic biology.

The goal of the fifth edition of Interlab is to discover the sources of variability in measurements and be able to correct them, so the measurements taken in different labs will not be variable anymore.

This year question is: Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or CFUs instead of OD?


Materials and Methods

Before the starting the experimental part, a plate reader was needed. Due to the lack of plate reader in our laboratory, it was kindly asked to Proteomics and Protein Chemistry Unit (DCEXS-UPF, PRBB) to use their equipment. The plate reader is the Synergy HTX Multimode Reader from Biotek, it can measure both absorbance and fluorescence. It has pathlength correction, which was not disconnected. It has control over temperature and it was set as room temperature (around 24-25 Celsius degrees). The excitation filter was 485/20 nm and the emission filter 528/20 nm and bottom optics were used. Moreover, the plates were black and flat-bottomed.

Eight plasmids need to be characterized in DH5-alpha E.coli strain in order to follow the protocol. The strain was obtained by collaboration with BIO-IQS iGEM team. The plasmids are the following: BBa_R0040, BBa_I20270, BBa_J3604000, BBA_J364001, BBa_J364002, BBa_J364007, BBa_J364008, BBa_J364009.

The materials used over the protocol are the same ones specified in the iGEM 2018 Interlab Study Protocol.

Results

Calibration

OD600 Reference Point


LUDOX CL-X

H 2 O

Replicate 1

0,056

0,034

Replicate 2

0,056

0,034

Replicate 3

0,056

0,034

Replicate 4

0,056

0,034

Arith. Mean

0,056

0,034

Corrected Abs600

0,022


Reference OD600

0,063


OD600/Abs600

2,864


Table 1 | Date from the OD600 Reference Point in calibration 1.

Particle Standard Curve


Fluorescence Standard Curve

This calibration needs to be repeated because of the low R 2 (R 2 =0,8547). But it can be caused by the plate reader since it was unable to read some of the biggest concentrations in the plate (10 uM of fluorescein).

Measurement

The measurement is still in process due to some problems with the transformation of the plasmids from the Parts Collection 2018.