A.3.1.4StudentResponseSheet
Activity 3.1.4 Student Response SheetPart I: DNA Microarray Virtual LabWhy are tissue samples from healthy and cancer cells taken from the same patient?It allows us to look at the differences in gene expression in cells that have the exact same genetic blueprint (accounting for disease and not differences in people).Describe the process used to isolate mRNA from the other types of RNA.Once the RNA is separated from proteins, DNA, and other cellular components, wash the RNA samples over columns filled with small beads, which only bind to mRNA. The other types of RNA will wash away, leaving only mRNA.Why is it necessary to make a cDNA copy? Why is mRNA not used?A fluorescent label (markers) in the cDNA molecule allows us to visualize the cDNA later on. mRNA is not used because it is less stable than DNA.Draw and label a diagram of the process of how labeled DNA copies are made. Your diagram should be in color.85725190500What happens once you apply the DNA from the two samples to the DNA microarray?They pair up with their complementary DNA strands that are on the microarray.What does the red color indicate?The red color indicates genes that are “turned up” in cancer (genes that produce more mRNA in the cancer cells than in healthy cells) (cancer cells).What does the green color indicate?The green color indicates genes that are “turned down” in cancer cells (healthy cells).What does the yellow color indicate?The yellow color indicates mRNA that is made in both healthy and cancer cells. It also indicates that the defects in cancer cells prevent the gene from being translated into a protein and cannot be detected using microarray analysis. (Therefore, it is probably not related to our study.)What conclusions can you make from microarray data?Microarray data analysis can determine which genes have been turned up, have been turned down, or are not affected (present in both cells). It can also determine whether or not the person expresses the trait.What are the limitations of DNA microarray technology?Defects in cancer cells that prevent the genes from being translated into proteins cannot be detected using microarray analysis (exactly which gene went bad to cause the cancer).Part II: Lung Cancer Microarray Wet Lab Use colored pencils or markers to draw what you observed in the microarray experiment.8572510223500104775161290Errors:Gene 2= purple, 1:10Errors:Gene 2= purple, 1:1Which gene(s) were expressed more in Grandpa Joe’s lung cells? How do you know?Genes 1 and 5: pink colored (cancerous)Which gene(s) were expressed less in Grandpa Joe’s lung cells? How do you know?Genes 3, 6: blue colored (normal/non-smoker)Were there any genes not expressed in either cell type? Explain why a gene would not be expressed in either cell.Gene 4Neither cell codes for that protein (unrelated, not coded for protein in lungs)Explain what it might mean for a gene to be expressed the same in both Grandpa Joe’s lung cell and a non-smoker’s lung cell.That gene is not influenced by smoking and doesn’t present much risk for cancer (codes for a protein they both need, like how to be a lung).Which genes may play a role in causing cancer in lung cells? Explain your choices.Genes 1, 5, 3, 6Dark pink/light pink colors and therefore are more expressed in cancer genesTurning up or down a protein-making process can cause cancerRecord your gene expression ratios for your microarray:Gene 1-- 8:1Gene 2—1:1Gene 3—1:8Gene 4—0:0Gene 5—2:1Gene 6—1:2 ................
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