Difference between revisions of "Team:Shanghaitech/InterLab"

Revision as of 18:59, 30 September 2017

233

Abstract

Repeatability is one of the most important part of biological experiment, however, it’s difficult to repeat the measurement data in different labs. As for the measurement of fluorescence, it can be influenced by various factors: bacterial strain, culture medium, plate reader and so on. Therefore, following a standard protocol strictly is one of the guarantee of the repeatable experimental conclusion. This year, we use plate reader rather than flow cytometry in 2016 (click here to view). Except for individual settings, we follow the standard protocol by iGEM authority completely, the data is reliable.

Plasmids and the prediction of results

All the eight plasmids are shown below:

From the table above, there are three different constitutive promotors (pcons) and two different ribosome binding sites (RBSs), which form six different combinations as Test Device 1-8.

(http://parts.igem.org/Part:BBa_J23101)

According to the data of iGEM parts, we know that the strength of these three pcons: J23101＞J23106＞J23117. However, there lacks the reliable data of the strength of pcon (J23151) and the comparison of three RBSs (B0032, B0034, J364100). In conclusion, we predict the result of the Interlab fluorescence measurement:

Test Device 1 ＞ Test Device 2 ＞ Test Device 3

Test Device 4 ＞ Test Device 5 ＞ Test Device 6

All the plasmid sequences (.dna file) can be download from: https://drive.google.com/file/d/0Bxb6U-RiSYsgQ1pTU0FsTG1SY1k/view?usp=sharing

Component cell strain & 96 well plate & Plate reader

Component cell strain

Brand: 康为世纪 (cwbiotech)

Strain: E.coli K-12 DH5α

96 well plate

Brand: Corning

-Black plate

-Flat-bottomed wells

Plate reader

Brand: BioTek

Instrument model: Cytation 5

Serial number: 1511021F

Pathlength correction: No

Number of flashes per well: 10

Orbital averaging (mm): 3.5

Fluorescence reading: top optic

Filter: Yes

Excitation wavelength (nm): 485/9

Emission wavelength (nm): 528/9

Protocol

(Some of the following steps are different from standard protocol from iGEM authority, the changes and the reasons are shown as red.)

Highlight

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Tooltips

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Dropcaps

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Blockquote

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Nulla nunc dui, tristique in semper vel, congue sed ligula. Nam dolor ligula, faucibus id sodales in, auctor fringilla libero. Pellentesque pellentesque eget tempor tellus. Fusce lacinia tempor malesuada. Ut lacus sapien, placerat a ornare nec, elementum sit amet felis. Maecenas pretium lorem hendrerit eros sagittis fermentum.

Fusce lacinia tempor malesuada. Ut lacus sapien, placerat a ornare nec, elementum sit amet felis. Maecenas pretium lorem hendrerit eros sagittis fermentum. Donec in ut odio libero, at vulputate urna. Nulla tristique mi a massa convallis cursus. Nulla eu mi magna. Etiam suscipit commodo ad gravida. Cras suscipit, quam vitae adipiscing faucibus, risus nibh laoreet odio, a porttitor metus eros ut enim. Morbi augue velit, tempus mattis sum dignissim nec, porta sed risus. Donec eget magna eu lorem tristique pellentesque eget eu dui. Fusce lacinia tempor malesuada. Ut lacus sapien, po anemat ornare nec, elementum sit amet felis. Maecenas pretium hendrerit fermentum lacus sapien, placerat a ornare nec fringilla libero convals.

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      <strong>Protocol</strong>
 </p>
-<p>
+<p class="MsoNormal"><span lang="EN-US">(Some of the following steps are different
-    (Some of the following steps are different from standard protocol from iGEM
+from standard protocol from iGEM authority, the changes and the reasons are
-    authority, the changes and the reasons are shown as</p><p color="red">red</p> red<p>.)
+shown as <span style="color:red">red</span>.)<o:p></o:p></span></p>
-</p>