RT-PCR A researcher wants to examine the effects of environmental pollutants pre
ID: 212214 • Letter: R
Question
RT-PCR A researcher wants to examine the effects of environmental pollutants present in the Calcasieu River on insulin receptor levels in skeletal muscle. The researcher gives one mouse water that was taken directly from the River and the second mouse receives tap water. After a month of drinking river water, the TA muscles of the mice are isolated and RNA is extracted. The RNA is converted into cDNA and RT-PCR is performed to look at insulin receptor levels. Answer the following questions. A. Why is he isolating RNA for these studies rather than DNA? (4 pts) (B.) Why must he convert the RNA into cDNA before performing RT-PCR? (4 pts) (C. Explain these results obtained àfter the RT-PCR and gel electrophoresis. (6 pts) 1 2 3 4 Lane 1-insulin receptor control rat Lane 2- insulin receptor experimental rat Lane 3- GAPDH control rat I ane 4-GAPDH stemid ratExplanation / Answer
A. We are studying about the expression of the receptor in this experiment. So, for gene expression we need to see the mRNA or protein profiles. DNA is always present in the genome it won't tell any thing abount gene expression that's why we are analysing the mRNA.
B. For quantification of RNA is difficult and also it degrade fastly. So we convert mRNA into cDNA by reverse transcriptase. In RT-PCR it provides the sufficient information about how much RNA expressed in the differernt experiments.
C. Lane 3 and 4 In this GADPH is a control or house keeping gene it always expressed in the cells.
Lane 1 - Control rat expressed normal amount of insulin receptor in the cell
Lane 2 - River water consuming rat shows decrease in the insuling receptor compare to the control normal tap water consuming rat.
It shows that pollutant river water affect the insulin receptor production in the cells.
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