Week Five: I thought growing cells was easy!

Although this was a shorter week for me as I had no work on Friday, I was still plenty busy.  After my successful PCRs last week, Pete and I selected a few samples that we would clone and clean-up for sequencing to see what was really being amplified in our samples.  Normally, this is a pretty easy process that just involves inserting the selected gene into "host" cells and then growing them on agar and picking colonies that grew successfully with said insert.  However, after plating some of these cells on Monday, Tuesday morning brought disappointment - barely anything grew!  This is very unusual as we normally have 20+ colonies, so I tried new cells and new vector (allows the gene to be inserted), but those plates didn't grow as well as usual.  This is something I'll be trying to troubleshoot in a few weeks when I do some more sequencing, but for now it worked well enough for my needs.

This process takes about 3 days total and I wasn't able to do much lab work as I am waiting on the results to see where to go from here.  Aside from reading papers and helping out with some various lab upkeep, I went sampling with Pete and his dog, Maggie, on Wednesday.  We went to Hammond Bay, Young's Bay, Ilwaco Harbor, and Willapa, taking one shore sample at each location.  We will be doing cell counts of these samples sometime next week and, if we see anything interesting, might do some PCR.  It was fun to get out of the lab for a day and go sample, although it was very windy and cold.

Finally, attached is my "midterm" presentation recapping the work I've done so far.  The map on the first slide shows our sample locations and the images on the second slide are from the DAPI/Proflavine/Euduboscquella probe staining we did of a Willapa sample.  It is a very good image of an infected ciliate, although this particular one is a tintinnid, not M. rubrum like we were hoping.

Until next week, Deirdre