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

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        <h3> Interlab </h3>
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<h3>★  ALERT! </h3>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2018.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2018.igem.org/Judging/Awards"> award listed below</a>. </p>
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2018.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
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        <div class="page-story">
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            <article id="Calibration" class="col-lg-10 col-lg-offset-1 col-md-12 col-md-offset-0 col-sm-offset-0 col-sm-12">
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                <header class="page-header">
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                <h1 id="model-introduction">Calibration experiments</h1>
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                </header>
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<p>Before starting cell measurements work, we accomplished three sets of calibration measurements with materials provided by the committee, including an OD600 reference point, a particle standard curve and a fluorenscein standard curve. These tests and data greatly facilitated our following quantitative interlab experiments.</p>
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                <h2>Calibration 1 OD600 reference point and LUDOX</h2>
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                <p>According to the protocol, we performed three calibration experiments (OD600 reference point, particle standard curve and the fluorescence standard curve) before the body part began.</p>
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<p>In this experiment, we first uesd LUDOX CL-X provided by the committee to obtain a conversion factor as the intermediate variable between Abs600 and OD600 measurement. The LUDOX solution (45% colloidal silica suspension) is hardly scattering and gives a low absorbance value, making it an ideal material for correct the standard path length of spectrophotometers in different laboratories, which would greatly improve the accuracy of the following volume dependent experiments.</p>
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                <p>Results shown below. </p>
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                <figure class="text-center">
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                    <img src="https://static.igem.org/mediawiki/2016/e/e9/Modeling_steps.png" alt="this is a pic"  width="60%">
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                    <figcaption>
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                        Table 1. OD600 reference point
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                    </figcaption>
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                </figure>
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                <p id="model-theory">4 replicates are measured respectively to acquire the average Abs600 of LUDOX CL-X solution and dH2O. Reference OD600 is given by the iGEM authority. The ratio of OD600/Abs600 is 3.652.</p>
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              <h2>Calibration 2 Particle Standard Curve</h2>
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                <p>After standardizing the absorbance value, we managed to conclude a conversion factor between Abs600 and the estimated number of cells. In this experiment, we used the silica beads delivered by iGEM committee and made a serial dilution in the 96-well plate according to the protocol. The beads’ size and optical charateristics are similar to cells and its concentration (amount of particles per volume)is known, allowing us to conduct a standard curve of particle concentration in 4 replicates. Origin data presented at the bottom of this page.</p>
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<figure class="text-center">
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                    <img src="https://static.igem.org/mediawiki/2016/e/e9/Modeling_steps.png" alt="this is a pic"  width="60%">
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                    <figcaption>
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                        Figure 1A Particle Standard Curve
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                    </figcaption>
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                </figure>
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<figure class="text-center">
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                    <img src="https://static.igem.org/mediawiki/2016/e/e9/Modeling_steps.png" alt="this is a pic"  width="60%">
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                    <figcaption>
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                        Figure 1B Particle Standard Curve (log scale)
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                    </figcaption>
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                </figure>
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<h2>Calibration 3 Fluorescence standard curve</h2>
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                <p>Fluorescence value is an important synthetic biology signal, but it can vary widely in different instruments. As a result, comparing the fluorescence output of test devices by creating a standard fluorescence curve can accurately standardize the amount and fluctuation of GFP fluorescencein in provided kits. In the fluorescence curve measurement, we used the fluorescein powder delivered by committee and made a serial dilution in the 96-well plate according to the protocol. 4 replicates were measured in total. Origin data presented at the bottom of this page.</p>
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<figure class="text-center">
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                    <img src="https://static.igem.org/mediawiki/2016/e/e9/Modeling_steps.png" alt="this is a pic"  width="60%">
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                    <figcaption>
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                        Figure 2A Fluorescein Standard Curve
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                    </figcaption>
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                </figure>
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<figure class="text-center">
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                    <img src="https://static.igem.org/mediawiki/2016/e/e9/Modeling_steps.png" alt="this is a pic"  width="60%">
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                    <figcaption>
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                        Figure 2B Fluorescein Standard Curve(log scale)
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                    </figcaption>
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                </figure>
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            </article>
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        <span>© 2018 BNU</span>
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<h1>InterLab</h1>
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<h3>Bronze Medal Criterion #4</h3>
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<p><b>Standard Tracks:</b> Participate in the Interlab Measurement Study and/or obtain new, high quality experimental characterization data for an existing BioBrick Part or Device and enter this information on that part's Main Page in the Registry. The part that you are characterizing must NOT be from a 2018 part number range.
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For teams participating in the <a href="https://2018.igem.org/Measurement/InterLab">InterLab study</a>, all work must be shown on this page.
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Revision as of 04:58, 11 October 2018

Team:BNU-CHINA - 2016.igem.org style = "font-family: Helvetica;"

Interlab

Calibration experiments

Before starting cell measurements work, we accomplished three sets of calibration measurements with materials provided by the committee, including an OD600 reference point, a particle standard curve and a fluorenscein standard curve. These tests and data greatly facilitated our following quantitative interlab experiments.

Calibration 1 OD600 reference point and LUDOX

According to the protocol, we performed three calibration experiments (OD600 reference point, particle standard curve and the fluorescence standard curve) before the body part began.

In this experiment, we first uesd LUDOX CL-X provided by the committee to obtain a conversion factor as the intermediate variable between Abs600 and OD600 measurement. The LUDOX solution (45% colloidal silica suspension) is hardly scattering and gives a low absorbance value, making it an ideal material for correct the standard path length of spectrophotometers in different laboratories, which would greatly improve the accuracy of the following volume dependent experiments.

Results shown below.

this is a pic
Table 1. OD600 reference point

4 replicates are measured respectively to acquire the average Abs600 of LUDOX CL-X solution and dH2O. Reference OD600 is given by the iGEM authority. The ratio of OD600/Abs600 is 3.652.

Calibration 2 Particle Standard Curve

After standardizing the absorbance value, we managed to conclude a conversion factor between Abs600 and the estimated number of cells. In this experiment, we used the silica beads delivered by iGEM committee and made a serial dilution in the 96-well plate according to the protocol. The beads’ size and optical charateristics are similar to cells and its concentration (amount of particles per volume)is known, allowing us to conduct a standard curve of particle concentration in 4 replicates. Origin data presented at the bottom of this page.

this is a pic
Figure 1A Particle Standard Curve
this is a pic
Figure 1B Particle Standard Curve (log scale)

Calibration 3 Fluorescence standard curve

Fluorescence value is an important synthetic biology signal, but it can vary widely in different instruments. As a result, comparing the fluorescence output of test devices by creating a standard fluorescence curve can accurately standardize the amount and fluctuation of GFP fluorescencein in provided kits. In the fluorescence curve measurement, we used the fluorescein powder delivered by committee and made a serial dilution in the 96-well plate according to the protocol. 4 replicates were measured in total. Origin data presented at the bottom of this page.

this is a pic
Figure 2A Fluorescein Standard Curve
this is a pic
Figure 2B Fluorescein Standard Curve(log scale)