happy spring break......

Monday we began by discussing our IJSEM, International Journal of Systematic and Evolutionary Microbiology, paper that we are turning in at the end of the month. We will be describing what we believe to be a new species belonging to one of the three genre we are focusing on: Geodermatophilus, Blastococcus, and Medestobacter. If we think we have strains with characteristics of a new genus we also have the option of describing those. We will describe their isolation and cultivation, colony morphology, chemotaxonomy, phylogenetic analysis, physiology, and taxonomic conclusions based on our research and data collected throughout the semester. In lab we plated out our desiccation spots by putting 500µl of liquid media into the wells and letting them sit for thirty minutes before mixing them with a pipette to resuspend the cells and spotting 10µl of the suspension onto a plate of its original media. We also made new stock plates of our 26 strains as well our soil samples, N97-6 and N97-19, that were positive when PCR was done with geo primers. We observed our 9kGy radiation plates. We scored strain 5 as ** and strain 25 and ****. The rest of the strains had no growth. The last thing we did was PCR with geo primers on our positive soil DNA that we extracted last week. Rainey made a premix with 80µl x10 buffer, 8µl geo primer 1, 8µl of geo primer 2, 80µl of dNTPs, and 632µl of water. 49.5µl of the premix was placed into each tube along with .5µl of DNA. We did a hot start to denature the DNA longer at 98C for 10 min. Next it was cooled to 90C and Rainey added .5µl of Taq since better results are achieved after adding the Taq later for these geo primers. Eugene did a wonderful job of running a gel of our results. Our soil DNA came out well. My groups 4126 isolate was positive, but not as bright as the others. On Wednesday we plated our second set of carbon sources:
C- control
F- fructose
CB- cellobiose
PY- pyruvate
TR- trehalose
SO/SB- sorbitol
MI- myo-inositol
MT- maltose
CT- citrate
AC- acetate
SC- succinate
MS- mannose
AL- alanine
AG- arginine
AS- asparagine
HS- histidine
LY- lysine
TH- threonine

We spotted 10µl of each strain onto agar containing each of the above. Finally, Lauren, Christi and Sean Michael purified PCR that had been done earlier in the day on some of our 4136 strains we do not have 16S for. Happy spring break everyone, have fun and be safe!!!!!