Our group is curious too! For this result we struggled to find an explanation for why no band appeared on the gel, but we pinned it on user error within our PCR which means we likely forgot to add one of the reagents, or we didn’t close the PCR tubes appropriately and that led to evaporation of our solvent in the thermocycler.
gDNA also known as genomic DNA is the natural DNA sequence including introns, exons, regulatory regions, etc. cDNA is generated in a lab, and is made by isolating the RNA translated from your genetic sequence and reverse transcribing it back into DNA. This produces a DNA sequence that is reflective of the RNA levels within the cell, and are efficient for measuring expression. The reason we generate cDNA rather than using RNA is because cDNA is much more stable.
Quantitative real time PCR is a technique for analyzing gene expression that is much more accurate and informative than the standard PCR we preformed in this experiment. Quantitative real time PCR provides a lot of information about the reaction as it proceeds, so we are able to analyze at exactly what point the gene is expressed, and it’s degree of expression using this technique.
curesymposium.org 
Great presentation! I’m curious to know what exactly happened with the cDNA/RPPO and why the PCR setup didn’t function well.
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Our group is curious too! For this result we struggled to find an explanation for why no band appeared on the gel, but we pinned it on user error within our PCR which means we likely forgot to add one of the reagents, or we didn’t close the PCR tubes appropriately and that led to evaporation of our solvent in the thermocycler.
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You talk about cDNA and gDNA in your results, what is the difference between these two? I did not quite understand the difference between these two.
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gDNA also known as genomic DNA is the natural DNA sequence including introns, exons, regulatory regions, etc. cDNA is generated in a lab, and is made by isolating the RNA translated from your genetic sequence and reverse transcribing it back into DNA. This produces a DNA sequence that is reflective of the RNA levels within the cell, and are efficient for measuring expression. The reason we generate cDNA rather than using RNA is because cDNA is much more stable.
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Can you expand more on what quantitative real time pcr exactly does?
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Quantitative real time PCR is a technique for analyzing gene expression that is much more accurate and informative than the standard PCR we preformed in this experiment. Quantitative real time PCR provides a lot of information about the reaction as it proceeds, so we are able to analyze at exactly what point the gene is expressed, and it’s degree of expression using this technique.
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