WOW we are officially back at school and doing this thesis thing!!
Life in the Williams Lab so far has consisted of lots of training.
I am doing my best to get the hang of working with the fish and learning the lab techniques so that I can (hopefully) start some juicy research soon!
I have read Larissa’s 2016 paper on Nfe2 as a regulator of the oxidative stress response during zebrafish development, as well as Quang’s thesis from last year. Both touch upon the project I will be undertaking, which involves me performing a gain of function assay to determine whether the downregulation of the Alas2 gene in Nfe2 KO fish is the (at least partial) cause of poor heme biosynthesis.
My meeting with Larissa today went great and we discussed my immediate goals, which include perfecting the PCR conditions for amplifying the Alas2 gene transcripts to make cDNA! Once I get this down pat, I can use TA cloning to put the cDNA in a plasmid, which will then be transformed into bacteria. From there, I will collect individual colonies, lyse the bacteria, get back the plasmid, and amplify it again, all so that I can inject the Alas2 gene back into zebrafish embryos to see if I can recover a normal heme phenotype. WOO!
For this week, in addition to fish breeding training, I’ve got to catch up on all the previous work that Quang did with PCR before he left. I’m also gonna read through some plasmid and TA cloning kit protocols.
Happy first full week! Catch ya on the flip side!