A. What is microarray? When would you use this experimental method? B. What is being differentially...
A. What is microarray? When would you use this experimental method?
B. What is being differentially labeled? How and why?
Homework Answers
A microarray is a molecular biology technique used to detect the expression of thousands of genes in the samples at particular time point. This allows the researcher to compare the gene expression of two different samples for example, healthy and cancer cells. DNA microarrays are chips that are printed with thousands of tiny spots known DNA sequence in defined positions. These known DNA sequence act as probes to detect the set of messenger RNA (mRNA) transcripts expressed by each sample. The whole set of mRNA in a cell is called as a transcriptome. Therefore, microarray is an important tool to study transcriptomics.
To perform a microarray analysis of two samples for example, healthy and cancer cells, following steps are required.
1. RNA is isolated from the samples
2. RNA is converted into complementary DNA (cDNA) by using reverse transcriptase PCR
3. Each cDNA sample is labeled with a fluorescent probe of a different color, for example, healthy cells are labelled with green fluorescent dye and cancer cells are labelled with red fluorescent dye.
4. Both the samples is mixed and allowed to hybridize with the probes on microarray chip.
5. If the expression of a particular gene is higher in the healthy cells than cancer cell, then the corresponding gene spot on the microarray appears green. Whereas, if the expression in the cancer sample is higher than in the healthy sample, then the spot appears red.
6. Based on this data a gene profile is created for healthy and cancer cells.
Please see the following diagram for
the summary
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A. What is microarray? When would you use this experimental
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