We use Escherichia coli as our chassis,the names of all strains are:Escherichia coli strain Nissle, BL21 Competent E. coli, DH5-Alpha Escherichia coli strain.Our chassis organisms are all widely-used E.coli strains.So they're all very safe, and even if they get out of the lab, the risk can be effectively controlled.
Difference between revisions of "Team:DLUT China/Safety"
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We wear plastic gloves, white coats and goggles when performing the experiment. In addition, the experimental team of our team conducted safety training and had a clear understanding and deep understanding of how to ensure safety during the experiment. During the experiment, we will use the knowledge we received during safety training to ensure safety.<br> | We wear plastic gloves, white coats and goggles when performing the experiment. In addition, the experimental team of our team conducted safety training and had a clear understanding and deep understanding of how to ensure safety during the experiment. During the experiment, we will use the knowledge we received during safety training to ensure safety.<br> | ||
| − | At the same time, we always guarantee the clean and orderly laboratory.<br> | + | At the same time, we always guarantee the clean and orderly laboratory.<br><br> |
<center><img src="https://static.igem.org/mediawiki/2018/5/50/T--DLUT_china--Lab1.jpeg" style="width:400px;height:380px;"/></center> | <center><img src="https://static.igem.org/mediawiki/2018/5/50/T--DLUT_china--Lab1.jpeg" style="width:400px;height:380px;"/></center> | ||
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Revision as of 15:13, 4 October 2018
Safety
Fluorescence is widely used as a representative of promoter activity. Although this is an indirect measurement, fluorescent proteins such as green fluorescent protein (GFP) provide useful insights into expression levels and can be continuously monitored without destroying the cells.
First, measure three calibration protocols: use LUDOX CL-X (45% colloidal silica suspension) as a single point reference to obtain conversion factor to convert absorbance (Abs600) to OD 600, use silica gel beads - microsphere suspension Obtain a particle standard curve,the fluorescein standard curve was measured with fluorescein at an excitation wavelength of 495 nm and an emission wavelength of this filter of 525 nm.
After making the standard curve, we successfully transformed the eight plasmids provided by the measurement committee and performed cell assays according to the protocol.
First, measure three calibration protocols: use LUDOX CL-X (45% colloidal silica suspension) as a single point reference to obtain conversion factor to convert absorbance (Abs600) to OD 600, use silica gel beads - microsphere suspension Obtain a particle standard curve,the fluorescein standard curve was measured with fluorescein at an excitation wavelength of 495 nm and an emission wavelength of this filter of 525 nm.
After making the standard curve, we successfully transformed the eight plasmids provided by the measurement committee and performed cell assays according to the protocol.
We wear plastic gloves, white coats and goggles when performing the experiment. In addition, the experimental team of our team conducted safety training and had a clear understanding and deep understanding of how to ensure safety during the experiment. During the experiment, we will use the knowledge we received during safety training to ensure safety.
At the same time, we always guarantee the clean and orderly laboratory.
At the same time, we always guarantee the clean and orderly laboratory.
Details
Discussion:
1. According to the Abs results at each time point 0 hours and 6 hours, the cell concentration of Device 1 is significantly lower than that of other devices.
2. According to the fluorescence raw readings, the fluorescence of Device 3 is weakest, the fluorescence of Device 4 is strongest. And the fluorescence of each group increased from 0h to 6 h, abnormally, the fluorescence of the two bacteria of Device 5 was different.
3. There is significant difference in plasmid conversion rate between negative control (device 1) and positive control (device 2).
2. According to the fluorescence raw readings, the fluorescence of Device 3 is weakest, the fluorescence of Device 4 is strongest. And the fluorescence of each group increased from 0h to 6 h, abnormally, the fluorescence of the two bacteria of Device 5 was different.
3. There is significant difference in plasmid conversion rate between negative control (device 1) and positive control (device 2).