Cultured Cells or Tissue 1. Pictured below are an ethidium bromide-stained agaro

Question

Cultured Cells or Tissue 1. Pictured below are an ethidium bromide-stained agarose gel (left) andac uyh tan a northern blot using an anti-sense probe for the human MMP-3 mRNA (right). (Note: ethidium bromide is a fluorescent dye that binds nucleic acids) The gels were both loaded with total cellular RNA from humanFiter yate in Comn tissue samples Prepare Lysate 1Lyse cells/homogenize tissue Spin 2 min 7, 14 Samples 1,8 2,93, 104,115,126, 1.3 Tissue Brain Heart Kidney Liver Lu M 1 2 3 4 5 6 ung Pancreas | Spleen Bind RNA Add ethanol, transfer and spin M 8 9 10 11 12 13 14 Wash Column Wash and spin 3 times MMP-3 Elute Total RNA Add elution buffer and spin 1 min Based on the data shown for the sample in question, how confident are you in the following statements? Explain how you came to your conclusion. What else could you say about the sample? Pure Total RNA A. There is no MMP-3 RNA in sample 1 B. There is no MMP-3 RNA in sample 13 C. There is MMP-3 RNA in sample 4 D. There is MMP-3 RNA in liver E. There is more RNA in sample 6 vs. sample 7 F. There is less RNA in sample 12 vs. sample 11 G. There is more MMP-3 RNA in spleen vs. kidney H. MMP-3 RNA is longer than most cellular RNA. I. MMP-3 RNA is shorter than 28S rRNA

Explanation / Answer

Human MMP3 mRNA is present in samples 1, 8, 4, 11, 5, 12, 7, 14. Samples 2, 9, 3, 10 also have the mRNA, however, in very little concentrations.

A.False

B. True

C. True

D. True

E. False. Sample 6 produced no band in northern blot. So it has no mRNA. Sample 7 is showing a band after blooting.

F. True. Sample 12 showed a lighter band compared to sample 11.

G. True. Spleen (sample 14) showed darkest band among all the other samples. Definitely, it contains higher amount of mRNA for MMP3.

H. True. In the electrophoresis gel, there are many bands below the corresponding MMP3 mRNA (corresponding estimated by looking at blot)

I. True (however, cannot be answered looking at the electrophoresis gel and blot, since no information is provided regarding 28S rRNA)

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