Which if the techniques (Southern blot, microarray-based hybridization, gel elec

Question

Which if the techniques (Southern blot, microarray-based hybridization, gel electrophoresis, PCR amplification, restriction enzyme digestion, Sanger-DNA-sequencing)

can be most appropriately applied to the experiments listed below.

Determine the size of a DNA fragment.

Isolate a specific DNA fragment coding for the insulin gene from pig given the DNA sequence information of the insulin gene.

Detect the existence of mutations in many disease causing loci in a patient in a high throughput way.

Determine the DNA sequence difference (ie. Detect SNP) between a piece of Chimp DNA and a homologous piece of human DNA.

Explanation / Answer

Answer:

1. Determine the size of a DNA fragment - Gel electrophoresis

Electrophoresis involves running a current through a gel containing the molecules of interest. Based on their size and charge, the molecules will travel through the gel in different directions or at different speeds, allowing them to be separated from one another. All DNA molecules have the same amount of charge per mass. Because of this, gel electrophoresis of DNA fragments separates them based on size only

2. Isolate a specific DNA fragment coding for the insulin gene from pig given the DNA sequence information of the insulin gene - Southern blot

A Southern blot is a method used in molecular biology for detection of a specific DNA sequence in DNA samples. Southern blotting combines transfer of electrophoresis-separated DNA fragments to a filter membrane and subsequent fragment detection by probe hybridization. A hybridization probe is a short (100-500bp), single stranded DNA. The probes are labeled with a marker so that they can be detected after hybridization.

3. Detect the existence of mutations in many disease causing loci in a patient in a high throughput way - Microarray-based hybridization

Microarray provides a basis to genotype thousands of different loci at a time, which is useful for association and linkage studies to isolate chromosomal regions related to a particular disease.

4. Determine the DNA sequence difference (ie. Detect SNP) between a piece of Chimp DNA and a homologous piece of human DNA - PCR amplification

Direct sequencing of conventional marker fragments led to the detection of allele-specific SNPs, and AS PCR was primed at these SNPs.

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