Chapter 101 (1/2)
Because of the particularity of prion, Zhou Wen was very careful when doing experiments.
Biological masks, biological gloves, biochemical protective clothing, fully armed.
Qin Haitao and the four of them just helped, and did not directly contact the vessels containing the virus.
There are four methods to detect prions.
1. Electron microscopy.
2, capillary SDS- gel electrophoresis.
3. Capillary electrophoresis immunoassay of fluorescent labeled peptide chain.
4. Two color intense fluorescence target scanning method.
, among them, the second ”gel electrophoresis” is the best and the most direct detection method.
PrPSc was purified by physicochemical methods such as differential centrifugation, ultracentrifugation, sodium n-lauroylsarcosine and PK.
was then performed capillary SDS- electrophoresis on a capillary electrophoresis apparatus.
The prion pathogen PrPSc was identified according to the time and shape of the absorption peak at 214nm.
This method needs less samples and does not need immunologic reagent.
The disadvantage is that we need to use ”ultracentrifuge” and ”special capillary electrophoresis instrument”.
At present, most of the ULTRACENTRIFUGES are purchased from foreign brands, and the price is as high as 500000 RMB.
And prion specific capillary electrophoresis instrument is also very expensive, requiring more than 200000 RMB.
Even if the price is high, there are few opportunities for key use.
General hospitals can't even earn money from purchasing equipment, and only the top three hospitals with money like Jiangzhou hospital will purchase.
Because of this, medical resources in big cities are scarce.
A strange disease in a small city, you can't even test it.
According to the physical and chemical methods, Zhou Wen prepared the reagent step by step carefully, extracted the diluent, and separated the variant prion protein on the ultracentrifuge.
Just upgraded biological immunology, let him more handy.
In this process, Zhou Wen also taught Qin Haitao and Dong Wenying about the points that should be paid attention to in the process of routine prion physical and chemical tests.
”When adding lauroyl, you must remember...”
These experiences, not to mention university professors, even those experts in prion research institutes in China, are not clear about them.
Because the domestic research on viruses, especially prions, is still in its infancy. Only the top foreign research institutes can master these practical experiences.
For example, the Institute of yeast prion, University of Kent, UK.
For a long time, the new discoveries in the field of prion proteins are almost monopolized by several yeast research teams.
Those top research institutes will not easily teach you technology.
Unless you get their offer.
Qin Haitao and Qin Haitao have not realized how lucky they are. They just think that Zhou Wen's explanation is very simple and easy to understand, and even tells them the essentials of his actions.
When Zhou Wen said that, Qin Haitao and Dong Wenying listened attentively.
As for the other two old experimenters, although they seemed to be listening, they actually didn't listen at all.
Because they think that it is impossible to extract prions by Zhou Wen's method, and such teaching is basically on paper.
Zhou Wen has no ”electric eye”. How can he know these specific experimental variables? Everything is just his guess.
After purification of PrPSc, Zhou Wen injected the supernatant containing prion into Beckman pace5500 for electrophoresis.
”OK ~” after the injection, Zhou Wen was relieved.
”Qin Haitao, you can stay and look after the machine, and others will wash their hands and eat.”
After hearing this, everyone went to the sink to wash their hands
After dinner, Zhou Wen didn't dare to go anywhere and went back to the laboratory directly.
The samples obtained by conventional extraction method are too few, so it's really uneasy not to look at them in person
At 7:30 p.m., the electrophoresis experiment was over.
In this experiment, one of the former experimenters thought that he would take a look at the failure of the experiment.
However, the next second, he showed a surprised look on his face, ”Hey, look, it seems that prion has been detected...”
On hearing this, several others gathered around.
”Well, that's true.”
”Teacher, come on, we have detected the prion.”
”Hiss It shouldn't be. How can it be? Is there a mistake... ”
Zhou Wen took over the analysis sheet and looked at it carefully, and soon his face showed irresistible joy.
No matter how well the previous experiments went, nothing was 100% until the results came out.For example, failure of capillary electrophoresis instrument can also lead to detection failure.