Why is agarose gel electrophoresis commonly used to analyse DNA fragments for verifying gene insertion?

Question

BlackTom AI · Accepted Answer

The question asks why agarose gel electrophoresis is commonly used to analyse DNA fragments for verifying gene insertion.
Option 1: 'It can directly visualise the expression of inserted genes in real-time.' This is incorrect because agarose gel electrophoresis is a DNA separation technique, not a real-time reporter of gene expression. Visualizing expression typically requires RNA or protein assays, such as RT-qPCR or Western blot, not a basic gel.
Option 2: 'It chemically modifies DNA fragments to make them fluorescent for easy detection.' In reality, agarose gels separate fragments by size and detection is achieved by staining intact DNA (e.g., with ethidium bromide or SYBR Safe) after separation, not by chemical modification of the fragments to become fluorescent. This option misstates the mechanism of detection.
Option 3: 'It separates DNA fragments based on size, allowing comparison with expected fragment lengths.' This is accurate. Agarose gel acts as a porous matrix; smaller fragments migrate faster than larger ones, so you can compare observed band sizes to the expected fragment lengths to verify whether a gene insertion produced the anticipated fragment sizes.
Option 4: 'It can accurately determine the nucleotide sequence of DNA fragments.' This is incorrect. Gel electrophoresis provides information about fragment size, not sequence. Determining sequence requires methods like Sanger sequencing or next-generation sequencing, not standard agarose gel analysis.
Thus, the correct rationale is that the method separates DNA by size, enabling checks against expected fragment lengths to confirm successful gene insertion. The other options describe capabilities (expression visualization, chemical modification for fluorescence, exact sequencing) that are not provided by simple agarose gel electrophoresis.

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Why is agarose gel electrophoresis commonly used to analyse DNA fragments for verifying gene insertion?

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