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5. (2 points) From the figure below, what can you conclude about the be sure to

ID: 186381 • Letter: 5

Question

5. (2 points) From the figure below, what can you conclude about the be sure to explain the significance of the beta-actin mRNA that is sh relatively consistent levels in all tissues. expression of PSAT mRNA and PHGDH mRNA? In your answer own. Note: beta-actin is a gene that is known to be expressed at Ii Figure 7 Northerm-bilot analysis of lissue distribution of PSAT-specitle MANA PSAT mRNA Muple-lissue Nothem-blot membranes (MINTM 1 and it Cion lech Labocalories, each containing 2 ug of poyAl)' RNAiane, wert hybiand withubeled PSATa, PHGDH a .andNAprobe. PHGDH MRNA P-actio mRNA

Explanation / Answer

According to the information, the Northern-blot analysis of PSAT mRNA has been performed in various organs. It was found that expression levels was highest in the testis followed by thymus. It was also found that the expression levels of PHGDH were also not uniform in these tissues. PHGDH is used as a loading control in these experiments to ensure that the correct amount of sample has been loaded in the samples thus confirming that there was no human error in loading the samples. This is because PHGDH is a house-keeping gene. However, un-even expression levels of this gene suggests that the loading was non-uniform.

This was further confirmed by the expression levels of beta actin. Beta actin, similarly, is a loading control. However, it is a cytoplasmic protein which is used as a loading control for cytoplasmic proteins. It can be clearly seen in the image that the expression levels of beta actin mRNA are also non-uniform in nature. This further confirms that the loading of samples was non-uniform and the expression levels of PSAT cannot be relied upon.

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