There are a few programs I used to design the primers. The main one was primer3plus.com which always you to input the sequence of your target gene and enter qualifications for your primers. The program then takes this information and selects prospective primer sequences that will bind to your gene.
Once you pick which sequence you want to use, you order the premiers of that sequence from a company that specializes in that type of production..
Hi Brooklyn,
In the case of Tetrahymena thermophila, since the cells undergo self induced dna damage during conjugation, if DNA damage did not occur, the resulting cells would likely be defective and dysfunctional. This could lead to apoptosis or a programed cell death pathway as well, but I would have to verify that claim.
To clarify, the SF label refers to the name of the primers used. So the SF bands show the PCR products from the PCR reactions containing either damaged or undamaged T. Thermophila mRNA and the designed SF primers.
To answer your question, the primers annealing to the DNA during PCR (polymerase chain reaction). During this process the sample dna and primers are combined and exposed to a repeated series of temperatures. During a period of high temperatures the DNA strands are denatured, causing them to separate from each other. Then when the temperature is brought down, the primers will bind to the DNA because they are complementary to specific segments on the the DNA strand.
So essentially, there isn’t a specific mechanical mechanism or tool used, it ends up happening because of complementary DNA sequences and being in close proximity to one another.
How did you go about designing the primers used?
Hey Connor,
There are a few programs I used to design the primers. The main one was primer3plus.com which always you to input the sequence of your target gene and enter qualifications for your primers. The program then takes this information and selects prospective primer sequences that will bind to your gene.
Once you pick which sequence you want to use, you order the premiers of that sequence from a company that specializes in that type of production..
Very good information! Regarding your information, if DNA Repair did not occur what would the effects be?
Hi Brooklyn,
In the case of Tetrahymena thermophila, since the cells undergo self induced dna damage during conjugation, if DNA damage did not occur, the resulting cells would likely be defective and dysfunctional. This could lead to apoptosis or a programed cell death pathway as well, but I would have to verify that claim.
As I am not very familiar with SF bands, may you please explain what that is and how it ties into how you got your results?
Hello Ella,
To clarify, the SF label refers to the name of the primers used. So the SF bands show the PCR products from the PCR reactions containing either damaged or undamaged T. Thermophila mRNA and the designed SF primers.
Hello! I really like this presentation. What lab techniques did you use to attach the primers? Thank you!
Hello Shae,
To answer your question, the primers annealing to the DNA during PCR (polymerase chain reaction). During this process the sample dna and primers are combined and exposed to a repeated series of temperatures. During a period of high temperatures the DNA strands are denatured, causing them to separate from each other. Then when the temperature is brought down, the primers will bind to the DNA because they are complementary to specific segments on the the DNA strand.
So essentially, there isn’t a specific mechanical mechanism or tool used, it ends up happening because of complementary DNA sequences and being in close proximity to one another.
What sort of research would need to be conducted to confirm that your gene is producing a protein that participates in DNA repair?
What is unknown about gene repair or in other words, what more could be learned to further the research?