my lab report!

ok since we're on the topic of sharing painful moments, I was thinking of explaining my latest lab report so that I can understand it better anyways. I'm kinda stuck, so explaining it out in layman terms would make me understand better. and it can teach you something too!

Alright, so basically the experiment involves us being provided with a gene of interest, and via in situ hybridisation (which I will explain later) we will create RNA probes that will be localised within particular organelles or tissues within the zebrafish embryo that contains the gene of interest.

You start off with being given a simple RNA sequence made up of only 4 nucleotides (AUCG) which runs off to about 200+ bases, which you can run off at the NCBI website.
I guess you could try inserting as many sequences made up of just A-C-U-G and they can link you up to a particular gene. use blastn. It's pretty cool.

ok back to in situ hybridisation. This involves a circular DNA plasmid that's just uh, circular, that contains the gene of interest. By linearising it and cutting it out using restriction enzymes, you can create complementary mRNA sequences. [For those with no bio backgrd, DNA is basically a whole recipe book -made up of only 4 bases A-T-C-G that codes for proteins all around your body. The mRNA is like specific recipes that are only taken out at particular events ie, if the cell needs it. The final dish would be the protein that the mRNA codes for, that makes up your skin, hair, signalling molecules, among all other proteins in your body.]These mRNA sequences are then mixed into a test tube containing zebrafish embryos in the hope that it will hybridise (stick to) the genes that it complements. The gene would only be found in organs that produces it and thus would highlight (using a coloured marker stuck to the mRNA) the organ. The final product would look something like this:



this is the right view of the embryo, highlighted using an mRNA that hybridises the keratin-8 gene which codes for skin proteins...



and this is a close up of the dorsal side of the embryo, where we can clearly see the muscle that has been highlighted by the myosin light chain 2.


And so being given the probe sequence, we're able to find out more about the gene and what it really codes for as found in scientific literature. And being given the pictures, we can deduce the morphological characteristics that it codes for specifically in this experiment (which took a total of 5 days! open lab prac, meaning we can go in anytime we're free to complete the experiment.)

And so now, knowing those two, the question is, so what? We're supposed to find out how much has been found in scientific literature about the development of the two genes, and infer our discovery from what has already been found. Either that or go back in time and pretend that we've found a novel discovery. Either way, we're required to write a ground break research paper in that format. So well, a lot of readings I have to do.

So, didn't understanding that make u feel a lot smarter? :)