Genetic Engineering Watch

OSAMARAJA
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#1
Report Thread starter 1 year ago
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For my coursework, I've been told to find an example of genetic screening involving PCR, Gel electrophoresis and DNA extraction.
Or to find a golden rice method that involves gel electrophoresis and PCR.
But I can't seem to find any
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QuentinM
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Report 1 year ago
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Ooo, i've done this!

For my first masters degree research project, i've been working with transgenic mice-mice with a genetic mutation in a particular gene of interest. However its not abundantly obvious when these babies are born which mice have it, and which dont (not like it stops eye formation or turns their skin a different colour). So we do the following:

1) Take a small clip of skin from their ear (sounds brutal on baby mice, i know. But could be much worse i guess)
2) Extract DNA from this tissue sample. because its genotyping and no-one cares that much about it, its normally done really crudely, by boiling the sample in mild bleach for a bit to break open the cells.
3) PCR the DNA with specific primers that will amplify a sequence in the gene of interest. You normally use three primers here, with 1 starter primer, and 2 primers to terminate. One terminates only in the transgenic mice, one only in the Wildtype mice (non-transgenic)
4) Gel electrophoresis is then used to separate the fragments amplified, which should be different sizes, and produce different bands on a gel.

So you could use mouse, or any animal, genotyping as an example. You take a tissue sample, extract DNA, PCR the gene of interest to see if its been mutated properly, then separate fragments to confirm the correct genotype. This process is pretty much done by any lab which handles mice and zebrafish, as no-way they dont deal with genetically modified mice in this day and age.
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