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Review Question - QID 106644

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QID 106644 (Type "106644" in App Search)
A scientist wants to determine if a specific fragment is contained within genome X. She uses a restriction enzyme to digest the genome into smaller fragments to run on an agarose gel, with the goal of separating the resulting fragments. A nitrocellulose blotting paper is then used to transfer the fragments from the agarose gel. A radiolabeled probe containing a complementary sequence to the fragment she is searching for is incubated with the blotting paper. Which of the following is the RNA equivalent of this technique?

Southern blot

10%

47/450

Northern blot

79%

354/450

Western blot

4%

20/450

qPCR

1%

5/450

RT-PCR

3%

14/450

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The technique used by the scientist to find a specific fragment in the genome was the Southern blot. The RNA equivalent of the technique is the Northern blot.

A Southern blot is used to examine for the presence of a particular DNA sequence with the use of a probe that is complementary to the sequence of interest. The first step is to digest the genomic DNA and separate it on an agarose gel. Due to the numerous bands, the gel will appear as a smear. The DNA on the gel is then denatured to create single strands. Next, the gel is then transferred onto a nitrocellulose blotting paper. Finally, a DNA probe, either radiolabeled or fluorescently labeled, which is complementary to the DNA region of interest, is added to detect the DNA fragment with the region of interest.

Incorrect Answers:
Answer 1: A Southern blot is used to analyze DNA to examine for the presence of a particular DNA sequence with the use of a probe that is complementary to the sequence of interest.
Answer 3: A Western blot is used to analyze protein expression.
Answer 4: qPCR is used to measure the gene expression. The mRNA is converted to cDNA, and a PCR reaction is performed with a probe that allows for quantification of gene expression.
Answer 5: RT-PCR is used to determine if a gene is expressed but not quantitatively. The mRNA is converted to cDNA, and a PCR reaction is performed and run on a gel to determine if the gene was expressed.

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