Team:BIT/Demonstrate
Demonstrate
As of 1 November, the tasks of the various sub-groups of the project are as follows:
Biosensor section:
What’s more, we will be going on all of our experiment to get better data and let our project more perfect and have a chance to apply for market.
Our first experiment is AFP and aptamer affinity verification test, this experiment is to determine whether our alpha-fetoprotein can capture the aptamer, if you can capture, then we can continue to do this project, Otherwise, the first step is to fail. Of course, we got positive results.
The second experimentally complementary strand was linked to lysine (Lys). This experiment plays a vital role in the process of transforming our macromolecule signals into small molecule signals. If there is no way to couple, then we have no way to make our engineering bacteria to identify and respond to the concentration of lysine, there is no way to complete the detection of alpha-fetoprotein.
The third part is the lysine deprotection experiment, trypsin can be targeted lysine, threonine and other amino acid residues in the peptide, but no literature clearly pointed out that trypsin can act on acid-modified lysine, so We decided to remove the protecting group to facilitate the subsequent digestion of trypsin.
The last experiment is the quantitative relationship between AFP and lysine. This experiment is a test of our experiments as a whole. Since it is very difficult to detect a lysine molecule, it is very clever in the course of the experiment. Method to verify the fluorescence intensity instead of lysine concentration, to complete the detection of alpha-fetoprotein.
Signal amplifier:
Our first experiment was to obtain lysine-deficient Escherichia coli, which was the only mechanism for determining the response of lysine by engineering bacteria to determine the effect of lysine concentration on bacterial growth.
The second experiment is the preparation of competent cells, in order to be able to better show the growth status of the strain, we need to enter the culture of bacteria to determine the amount of fluorescent expression, so the preparation of competent cells is essential step.
The third experiment is to study the growth status and fluorescence expression of engineering bacteria, divided into different concentrations, the same time and the same concentration, different time.
Finally, we connect the successful plasmid to freeze into dry powder and deliver it to iGEM official
Microfluid chip:
First of all, we carried out a double-chamber chip with gelatin and mask, and then we carried out a magnetic bead fixation experiment on the chip to ensure that the buffer did not punch the beads from the first chamber The second chamber.
After that, we conducted a light test on the chip, get a better result. Chip and laboratory experiments carried out in parallel, we measured the growth curve of the engineering bacteria and the determination of fluorescence expression, and achieved a better degree of consistency with the laboratory results.
Finally, we carried out freeze-drying and resuscitation experiments on the engineering bacteria to ensure that the engineering bacteria could survive in the absence of lysine for long-term survival in the chip and resume growth and expression in the event of a suitable condition.
IMPORTANT:
Microfluidic chip
1.Microfluidic chip production
2.Magnetic bead fixation experiment
3.Bacterial growth curve and fluorescence determination
4.Freeze - dried bacteria experiment
