I’ll try to demonstrate the extent of my understanding & hopefully any of you fellow experts can fill in the gaps for me
So we know that we run Gel Electrophoresis, the DNA gets separated into many fragments and are sorted according to their sizes. But……
1) WHAT caused it to separate in the first place? Is it the voltage that breaks the DNA into fragments? Or is it the restriction enzymes that cut at the restriction recognition sites? How does it get fragmented?
2) Why are some fragments larger/smaller than others? I don’t understand how there are many fragments that have the same size?
3) Why does each person’s DNA show different bands?
Besides the above, I am also trying to link between performing a PCR then Gel Electrophoresis for visualisation and Sanger sequencing. I understand that PCR amplifies the region that we’re interested in, and then we use Gel Electrophoresis to confirm that they were amplified and there are no mutations. What would we have seen on the Gel if there had been mutations? What does sequencing do?
Would appreciate some feedback so we can discuss in the comments x
In: Technology
>What would we have seen on the Gel if there had been mutations?
Depends entirely on the kind of mutation. A large enough insertion/deletion will be visible after running a gel because the larger-/smaller-than-normal fragment will be less far/further along the gel than a wild type band.
SNPs won’t be visible through gel electrophorersis.
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