A PCR test is a polymerase chain reaction, which is used to generate massive amounts of DNA from a small DNA sample by exponentially replicating the DNA. PCR is used in this experiment because the expression of cDNA (that was reverse-transcribed from Rrm2 mRNA) is measured, and it is much easier to measure this expression if a large quantity of this cDNA is present.
Upregulated means that a gene is expressed more during a certain condition. In this experiment, Rrm2 is upregulated, because it increases in expresion, when DNA damage occurs in T. Thermophila.
The GS primers were the primers that only annealed to the Rrm2 gene, and could then be used to amplify Rrm2 cDNA and determine if Rrm2 expression increases during DNA damage.
The control primers were primers that annealed to control genes that confirmed that the experiment worked correctly. These primers included: Atpv0c6 primers that showed that the PCR was run correctly and also showed that a correct amount of cDNA was added to each lane. Rad51 primers were also used to amplify the Rad51 gene, to show that DNA damage did occur.
What is a PCR test and why was it utilized in these experiments? Thanks!
A PCR test is a polymerase chain reaction, which is used to generate massive amounts of DNA from a small DNA sample by exponentially replicating the DNA. PCR is used in this experiment because the expression of cDNA (that was reverse-transcribed from Rrm2 mRNA) is measured, and it is much easier to measure this expression if a large quantity of this cDNA is present.
Hi Nick!
Can you please expand on what “upregulated” might mean?
Thank you!
Avalon
Upregulated means that a gene is expressed more during a certain condition. In this experiment, Rrm2 is upregulated, because it increases in expresion, when DNA damage occurs in T. Thermophila.
What are the GS and control primers that was used to amplify the cDNA?
The GS primers were the primers that only annealed to the Rrm2 gene, and could then be used to amplify Rrm2 cDNA and determine if Rrm2 expression increases during DNA damage.
The control primers were primers that annealed to control genes that confirmed that the experiment worked correctly. These primers included: Atpv0c6 primers that showed that the PCR was run correctly and also showed that a correct amount of cDNA was added to each lane. Rad51 primers were also used to amplify the Rad51 gene, to show that DNA damage did occur.
What would happen if the genes for DNA repair are damaged or are otherwise nonfunctional