The (relatively) decent status of my research
The work is actually going pretty well! What I’ve done of importance:
Isolated and run rtPCR on the first of three RNA samples from the neural mutant and wild type strains of C. elegans, yielding (surprise!) insubstantial results. However, last Friday I isolated RNA from the second samples of C. elegans, and we’ll run these soon to determine the immune gene expression levels in both strains.
The results from the first run may have been messy because of a possible screw-up by Katie in the mixing of samples in the first run, so I’ve been doing PCR and gel runs to determine if two of the samples were mixed. We used a primer we thought was specific for the experimental pathogenic species, and would then see if that primer amplified any of the DNA in the E.coli samples–if so, then the contamination would be confirmed. However,the primers were (unfortunately) not specific for the pathogen; in fact, they amplified DNA in all the samples we had, and then after I did another PCR and gel run, we determined that the primers definitely amplified DNA from the C. elegans, E. coli and pathogen. So we’re just waiting for the next results of rtPCR to compare them to the original, and will do a third and, for me, final RNA isolation and analysis to determine with some certainty the changes in gene expression.
I’m also choosing the best couple of transgenic mutants that produce green flourescence when their immune gene is expressed, crossing them with our mutant strain and will compare their levels of flourescence on pathogen and E.coli. No green flourescence, no gene production!
In other news, it’s the season of prelims in the MGM department, so we’re celebrating two of our grad students’ passing this week! Congrats to Ine and Mickey!