Difference between revisions of "Team:HUST-China/InterLab"
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<li class="active"><a href="#section1">Introduction</a></li> | <li class="active"><a href="#section1">Introduction</a></li> | ||
<li><a href="#section2">Provenance and Release</a></li> | <li><a href="#section2">Provenance and Release</a></li> | ||
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| − | + | <li>① Prepare your 96 well plate</li> | |
| − | + | <li>②Add 100 μl LUDOX 100 % into wells A1, B1, C1, D1</li> | |
| − | + | <li>③Add 100 μl of dH2O into A2, B2, C2, D2</li> | |
| − | + | <li>④Measure absorbance 600 nm of all samples in all standard measurement modes in instrument</li> | |
| − | + | <li>⑤Import data into "Abs600" blue cells in provided Excel calibration sheet</li> | |
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| − | + | <li>① Spin down fluorescein stock tube to make sure pellet is at the bottom of tube.</li> | |
| − | + | <li>② Prepare 2x fluorescein stock solution (100 μM) by resuspending fluorescein in 1ml of 1xPBS</li> | |
| − | + | <li>③ Dilute the 2x fluorescein stock solution using 1xPBS to make a 1x fluorescein solution (final concentration is 50 μM).</li> | |
| − | + | <li>④ *Illustration of serial dilution samples in 96 well plate or cuvettes: value decreases by 2-fold with each column (50% in column 2, 25% in column 3, 12.5% in column 4, etc.)</li> | |
| − | + | <li>⑤Import data into "Abs600" blue cells in provided Excel calibration sheet</li> | |
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Revision as of 08:20, 16 October 2017
「Interlab」
Introduction
It is significant for synthetic biology to develop a reliable and repeatable measurement, the same to all the other engineering disciplines. We HUST-China have volunteered to test some RBS devices (BCDs) that are intended to make gene expression more precise and reliable by measure the expression level of GFP, in order to help the iGEM community collect data about how reliable will these devices turn out to be in labs around the world.
Provenance and Release
①Individuals responsible for conducting InterLab study
| Individuals | Interlab Part |
|---|---|
| Kangyuan Yu, Haibo Huang, Ziyang Xiao, Shaofeng Liao | created the devices |
| Efan Wang, Long Cheng, HuiPing Shi | conducted the measurements |
| Efan Wang | processed the data |
②Corresponding email
| Efan Wang | erfan@hust.edu.cn |
|---|---|
| Kangyuan Yu | 985930862@qq.com |
| Haibo Huang | u201512127@hust.edu.cn |
| Ziyang Xiao | 372657289@qq.com |
| Shaofeng Liao | 15827233830@qq.com |
| HuiPing Shi | 172295915@qq.com |
| Long Cheng | u201512127@hust.edu.cn |
Chassis and Safety Information
What chassis did you use?
Escherichia coli DH5alpha
What Biosafety Level is your chassis?
BSL1
What PPE did you utilize during your experiments?
Tianming gloves
Songxinjiujiu labcoats
Instrument Information
hat instrument did you use during your measurements?
plate reader
Please provide the brand and model of your instrument.
Flexstation 3
Calibration Protocol
A1. Protocol for Optical Density (OD600) Standard Measurement
Did you use pathlength correction during measurement?
Yes
Number of flashes per well
6
Orbital averaging (mm)
600
What temperature setting did you use during the measurement?
22℃
What type of 96-well plate did you use?
Black plate (preferred)
Did your plate have flat-bottomed or round-bottomed wells?
Flat
A2. Measurement Steps
B1. Protocol for FIuorescein Fluoresence standard curve
Did you use pathlength correction during measurement?
Yes
Number of flashes per well
6
What gain setting did you use?
Automatic
If you used a filter, what light wavelengths did it pass?
530nm
Emission wavelength
530nm
Excitation wavelength
485nm
Fluorescence reading
Bottom optic
What type of 96-well plate did you use?
Black plate (preferred)
Did your plate have flat-bottomed or round-bottomed wells?
Flat
What temperature setting did you use during the measurement?
22℃
