Genome Last Quiz Flashcards
What was the purpose of the first PCR done in the experiment?
The purpose of the first PCR was to determine the quality of the RNA.
What was the purpose of the Actin PCR?
The purpose of the Actin PCR was to determine whether or not cDNA was present. If cDNA is present, it tells us that the RNA is present which will allow us to test for the expression of genes.
How many strip tubes will be used for a PCR contains X reactions?
X-1
What was the purpose of the last PCR?
The purpose of the final PCR was to run the gene-specific primers; through doing this, you would see whether or not your assigned gene was expressed in the respective tissue. If there was an appearance of a band, it tells us that the gene was expressed in that certain tissue.
How do you know how much of the Master Mix to aliquot per strip tube?
Divide the total required by the number of strip tubes present.
If no bands showed up in the gene-specific PCR, what does that mean?
The absence of bands means that the assigned gene was not expressed in the particular tissue.
If multiple bands showed up in the gene-specific PCR, what can it be attributed to?
Non-Specific primer binding and the contamination of gDNA. When it comes to non-specific primer binding, there might have been regions in the cDNA similar to the region that intends to be amplified. When it comes to gDNA contamination, the presence of gDNA would lead to the presence of multiple bands (intron and exon combo).
What is the equation for balancing concentration and volume?
C1V1=C2V2. C1= Stock concentration; V1:= stock volume. C2= final concentration ; V2= final volume.
