Gregory Nugent's blog

So, this week was interupted with me catching a terrible cold at the tail end of it.  Thus I am finishing everything a little later (aka the following Tuesday).  However, the last LacZ assay has been run, and...  INCONCLUSIVE!  Fun stuff and I am not even all that surprised anymore.  So yeah, the resD mutant strains seem to be odd little buggers who do not allow me to gather clean and repeatable results.  Instead of verifying one or the other of the two results, all I did was confirm that they both were viable results, grr!  So, what does this all mean ladies and gents?  That some

This week definitely had its ups and downs, but I am going to focus on the positives J. I finally have had all four of the plasmids I’ve been constructing come back clean from sequencing! On Monday I had received the news that once again the plasmid pFS1, which was prepared by the highschool intern Felicia, had yet again failed to come back clean from sequencing. Dejected, since I really was hoping I would be successful this time and begin the transformation into Bacillus subtilis with all of my plasmids.   So, we decided to reculture and reconcentrate at a higher level the pFS1-1 st

This week definitely had its ups and downs, but I am going to focus on the positives J.  I finally have had all four of the plasmids I’ve been constructing come back clean from sequencing!  On Monday I had received the news that once again the plasmid pFS1, which was prepared by the highschool intern Felicia, had yet again failed to come back clean from sequencing.  Dejected, since I really was hoping I would be successful this time and begin the transformation into Bacillus subtilis with all of my plasmids.   So, we decided to reculture and reconcentrate at a higher level the pFS1-1

This week has been a mixed bag.  It started out well.  I got to begin by learning how to transform the Bacillus subtilis and by inserting my whole promoter sequence (which came back successfully two weeks ago from sequencing) into the wild type strain of competent cells I generated last week.  This involved making DSM agar with Erythromycin as the antibiotic resistance selection agent (since the plasmid will give this resistance to the B.subtilis which it infects).  When I came back on Tuesday, however, I arrived to the unpleasant surprise of having colony growth on my con

I had no mistakes which needed redoing this week! That was a super plus. Also, I amazingly got back my sequence by Monday morning (we didn’t think we would be getting it till like Thursday), and it showed that I had a perfect plasmid (not the case with Keifer – the high school teacher). So for me, that meant I am ready to transform this plasmid into Bacillus subtilus next week.  Before I can do that, however, I needed to make my B.

Yay!  This week went fairly smoothly and without any major surprises (well, for the most part...).  I spent Monday redoing about three days worth of work from week one, all in one day.  Very frustrating that I had to do it, but great practice and I thankfully was successful this time at creating the expression plasmid.  The rest of the week in the lab centered around creating a competent E. coli colony and transfecting into it my first expression plasmid for replication.  This also involved growing the E.