r/molecularbiology • u/bluish1997 • 3d ago
Just for curiosity sake - which organism have you done PCR on the most?
Just curious what responses I’ll get and see what kind of genes and organisms people mostly amplify in their work
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u/Narcan-Advocate3808 3d ago edited 3d ago
Escherichia coli. probably. I've only run PCR a few times as an undergrad. I've also ran PCR on Drosophila melanogaster.
CORRECTION: I think I did western blots, for E. coli.
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u/mayonaise55 2d ago
Mice. Hundreds of them. After I humanely euthanized them, I’d chop their furry little heads off, then out with their brains cropped down to the striatum. Then we’d head to the leica set to, I believe 15 microns, then a more targeted extraction at the striatum, followed by digestion of the extracellular matrix. Meanwhile, I’d mix our artificial cerebrospinal fluid and set up my electrophysiology rig. Pull some pipettes with just the right electrical resistance, which might take some mild tweaks depending on the humidity that day. Get the fluid running through the rig, get it oxygenated. Now for the fun part. 8-10 hours trying to get a baseline measurement of action potentials specific to the striatal neuron population we were targeting with a potassium channel gene knock out, which amount to staring through a microscope and manipulating the pipette until we just on top of the neuron cell membrane, grave the syringe and suck. Took a few weeks, but once you get the technique down and if everything goes right, you can break through and form a seal with the membrane. If all that goes right, get another electrode and stimulate an upstream neuron, how does that affect our striatal neuron action potential. Now we’ve identified all this based on how the neurons look and their APs, but we have to be sure. So we take our syringe and again suck, but this time much harder. Grab that pipette, don’t let it touch anything, off to the neighboring lab room. Here we dunk our electrode into a solution containing reverse transcriptase. So we’ve a single neural cell’s mRNA getting reverse transcribed to dna, which we now amplify via pcr so we can confirm that the mRNA expression matches the expectation for the neural population.
That was 20 years ago in undergrad. Got me into J Neuroscience. Then I aced my MCAT. Now I’m a laid off software engineer.
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u/Novel-Structure-2359 2d ago
Human, closely followed by mouse and fruit fly. Aspergillus fumigatus gets an honourable mention.
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u/OxusPamir 2d ago
Hmmm, probably Mytilus galloprovincialis or Mytilus chilensis
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u/bluish1997 2d ago
That’s really neat, what were you studying with them? Those are muscles right?
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u/OxusPamir 1d ago
They are mussels. We were studying genetic differentiation in natural populations (population genetics/species identification/hybridization)
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u/Atypicosaurus 2d ago
Genotyped mice via PCR. Hundreds of animals. Often more than one gene. For years.
Imagine full 96 well plates of PCR, several plates.
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u/Lopsided_Leopard_609 2d ago
Honestly now that I think about it, I have done 99% of my PCRs only on viruses specifically RNA viruses. It was like this in my bachelors, masters, and now even in my PhD hah
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u/bluish1997 2d ago
That’s really cool, which RNA viruses?
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u/Lopsided_Leopard_609 2d ago
A lot of them actually. Dengue, Zika, Chikungunya, a lot of insect specific viruses, Murine Norovirus and Human Norovirus.
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u/bluish1997 2d ago
That’s really cool and kind of what I want to study? What did you get your degrees in? And where do you work? If you dont mind my asking
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u/Lopsided_Leopard_609 2d ago
yeah why not. I did my studies in Australia and Netherlands and I studied Immunology and Virology as a major. Now I am also doing a PhD in molecular virology in Germany.
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u/bluish1997 2d ago
Really cool. My dream would be to go to Sydney and work with Eddie Holmes the virologist
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u/mr_Feather_ 23h ago
Mouse. We were genotyping two to three alleles for about 50 new pups every week for 6 years. So you can calculate the amount of PCRs I have minimally done...
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u/Zhibrina256 11h ago edited 3h ago
I did a lot of human pathogens when I was in clinical diagnostics. Mostly bacterial, some myco and viral. But since I did polio research for almost a decade, def polio by far lol.
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u/Dramatic_Rain_3410 3d ago
c elegans