At the conclusion of our lab we found that two people in our class were infected with a "disease" that they had unknowingly spread to their classmates after exchanging "bodily fluids". In our lab we began by taking a sample from everyone (these were pre-made by Mr. Chugh) and exchanging them by gently mixing the samples with three people besides ourselves. When we came back the next day after storing the samples at 4 degrees celsius overnight, the found each of our samples.
We loaded each sample into two wells and then added two wells of each positive and negative control to compare our results to. After waitng four minutes we were able to "wash" the samples out with a wash buffer.Then we transferred 50 microliters of antibodies into each well so that the antigens could bind to them. After waiting 4 minutes we washed the samples two more times. After the washing we added 50 microliters of secondary antibody and then waited 4 minutes again, following with three more washes. Lastly we added 50 microliters of enzyme substrate into the twelve wells, this made the contaminated samples turn blue. At our lab table both Elizabeth's and mine were a strong blue, while Ayan and Nate were not contaminated. After surveying the class's results we were able to determine that Taylor Grey and Chloe Krey were the original holders of the "disease.
Sources of error: I shared with four people. My last person being Michael who had just shared with Chloe, leading to my contamination!
Monday, May 23, 2011
Monday, May 16, 2011
Sharing Too Much
In this lab we will be sharing "bodily fluids" with our classmates. We will use a process called ELISA to determine if we have been infected with a contagious disease. ELISA (Enzyme-linked Immunosorbent Assay) is used to detect the presence of a disease agent. When a person is exposed to a "disease agent" their body will react with an immune response. The molecules it uses are called anitgens, within a couple days antibodies ( proteins that recognize the anitgen and bind very tightly) begin circulating in one's body. Anitobodies are made when the body has an immune response.The anitgens are created and then after a period of time the antibodies will begin to specicficallt recognize the anitgen.
The process called ELISA is used for many tests, including pregnancy tests, disease detection, drug testing, testing indoor air quality, and determining if food is labeled correctly. ELISA is used to easily and quickly detect whether patients have been exposed to certain viruses of conditions.
ELISA has four steps:
1. The anitgen is added to the wells of the microplate strip and incubated.
Unbound antigen is washed from the wells with a detergent.
2. Primary anitbody solution is added to the wells and incubated to allow antibody to bind to the antigen.
3. Enzyme- labeled secondary antibody solution is added to the wells.
4. Chromogenic enzyme is added to the wells and incubated to allow color to develope. Then the results are looked at, wells that are colorless are negative and wells that turn blue are positive.
The process called ELISA is used for many tests, including pregnancy tests, disease detection, drug testing, testing indoor air quality, and determining if food is labeled correctly. ELISA is used to easily and quickly detect whether patients have been exposed to certain viruses of conditions.
ELISA has four steps:
1. The anitgen is added to the wells of the microplate strip and incubated.
Unbound antigen is washed from the wells with a detergent.
2. Primary anitbody solution is added to the wells and incubated to allow antibody to bind to the antigen.
3. Enzyme- labeled secondary antibody solution is added to the wells.
4. Chromogenic enzyme is added to the wells and incubated to allow color to develope. Then the results are looked at, wells that are colorless are negative and wells that turn blue are positive.
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