Difference between revisions of "Team:WHU-China/InterLab"

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             <li id="nv0" style="width:56px"><a href="#">logo</a></li>
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             <li id="nv0" style="width:200px"><a href="https://2017.igem.org/Team:WHU-China"></a></li>
 
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               <a>Project</a>
 
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                 <li id="nv11"><a href="#">Description</a></li>
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                 <li id="nv11"><a href="https://2017.igem.org/Team:WHU-China/Overview">Overview</a></li>
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                 <li id="nv12"><a href="#">Design</a></li>
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                 <li id="nv12"><a href="https://2017.igem.org/Team:WHU-China/Background">Background</a></li>
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                 <li id="nv13"><a href="#">Experiments</a></li>
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                 <li id="nv13"><a href="https://2017.igem.org/Team:WHU-China/Introduction">Introduction</a></li>
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                 <li id="nv14"><a href="#">Results</a></li>
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                 <li id="nv14"><a href="https://2017.igem.org/Team:WHU-China/Designs">Designs</a></li>
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                <li id="nv15"><a href="https://2017.igem.org/Team:WHU-China/Experiments">Experiments</a></li>
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                <li id="nv16"><a href="https://2017.igem.org/Team:WHU-China/Results">Results</a></li>
 
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             <li id="nv2"><a>Parts</a>
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             <li id="nv3"><a href="#">Interlab</a></li>
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             <li id="nv3"><a>Interlab</a>
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             <li id="nv4"><a href="#">Models</a></li>
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                <li id="nv31"><a href="https://2017.igem.org/Team:WHU-China/Interlab">Results</a></li>
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                <li id="nv32"><a href="https://2017.igem.org/Team:WHU-China/Calibration">Calibration</a></li>
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            <li id="nv51"><a href="https://2017.igem.org/Team:WHU-China/Model">Modeling</a></li>
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                 <li id="nv81"><a href="https://2017.igem.org/Team:WHU-China/Member">Member</a></li>
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                 <li id="nv83"><a href="https://2017.igem.org/Team:WHU-China/Collaborations">Collaborations</a></li>
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<h6>Description</h6>
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  <li><a href="#Brief">1 Brief introduction in RdhANP</a></li>
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  <li><a href="#Description">2 Description of our operating bacteria</a></li>
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        <h1>Interlab Results</h1>
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  <h3>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;GFP is one of the most used markers in synthetic biology. It gives researchers an access to continuously monitoring the expression level of specific plasmid. However, due to variation in units, methods of data processing, protocols, or instruments, it is hard to repeat measurement in different labs. To solve this problem, we participate in interlab study by quantifying the experiment with standard protocol.</h3>
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<h3>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Lack of black wells with transparent bottom, we had to use black wells with black bottom in fluorescence measurement and transparent wells with transparent bottom in Abs measurement. We strictly followed CALIBRATION PROTOCOL and CELL MEASUREMENT PROTOCOL. You can see our experimental records in NOTEBOOK_WET LAB</h3>
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<p align="center"> <img src="https://static.igem.org/mediawiki/2017/1/17/WHU-China-fluor1.png"  alt=""/></p>
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<p align="center"><img src="https://static.igem.org/mediawiki/2017/d/d9/WHU-China-fluor2.png"  alt=""/></p>
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<h3>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;After analyzing our data and discussing with iGEMers from Team-ZJU and Team-FUFA, we strikingly found that the value of our OD600/Abs600 is as high as 3.37972167. Many reasons might account for it, such as instruments of varies brands and hours of use, and detailed experiment process done by different people and in different lab environment. </h3>
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<h3>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;As we can see from the above two figures, values nearly form a straight line on both linear and log scale and slope of their trend lines is 1:1, which means we didn’t have consistent pipetting error. But our coefficient of determination(R2) of both lines can’t reach 1, so there’re still lots of work for us to figure out the deviation. We hope that our data can do some help to other Teams or researchers in solving the problem of repeating measurement in different labs.</h3>
  
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          <h1 id="Brief">Brief introduction in RdhANP</h1>
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          <p>&nbsp;&nbsp;&nbsp;&nbsp;RdhANP is a native protein of Nitratireducror pacificus pht-3B. It is codon-optimized for operated and expression in Bacillus magaterium. RdhANP can catalyze the reduction of ortho-halogenated phenolic compound, which endows it with biological dehalogenation function. Different from other reductive proteins that are membrane-associated and oxygen-sensitive in their family, RdhANP is a soluble cytoplasmic one and oxygen-tolerant, which is fairly good for heterologous expression in other chassis. However, this enzyme displays a strict requirement for o-halogenated phenolic substrates and B12 cofactor; it also should under reductive condition using either reduced methyl viologen or NADPH. </p>
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Revision as of 00:50, 31 October 2017

Interlab Results

         GFP is one of the most used markers in synthetic biology. It gives researchers an access to continuously monitoring the expression level of specific plasmid. However, due to variation in units, methods of data processing, protocols, or instruments, it is hard to repeat measurement in different labs. To solve this problem, we participate in interlab study by quantifying the experiment with standard protocol.

         Lack of black wells with transparent bottom, we had to use black wells with black bottom in fluorescence measurement and transparent wells with transparent bottom in Abs measurement. We strictly followed CALIBRATION PROTOCOL and CELL MEASUREMENT PROTOCOL. You can see our experimental records in NOTEBOOK_WET LAB

         After analyzing our data and discussing with iGEMers from Team-ZJU and Team-FUFA, we strikingly found that the value of our OD600/Abs600 is as high as 3.37972167. Many reasons might account for it, such as instruments of varies brands and hours of use, and detailed experiment process done by different people and in different lab environment.

         As we can see from the above two figures, values nearly form a straight line on both linear and log scale and slope of their trend lines is 1:1, which means we didn’t have consistent pipetting error. But our coefficient of determination(R2) of both lines can’t reach 1, so there’re still lots of work for us to figure out the deviation. We hope that our data can do some help to other Teams or researchers in solving the problem of repeating measurement in different labs.