Priming the ISH

I am really lacking creativity at the moment so I apologize for the pathetic title.

Today we had a full day planning our ISHs.  The first step was making sure that our primers worked, which was especially important considering we had several new primer sets to test out for a variety of DNA:  Three primers for our putative pathogen for Sea Star Wasting Disease and a suite more to try and figure out possible Rickettsia-Like Organisms (RLOs) in coral.  We tested this by running a large number of PCRs – five different cycles – to double check that our primers amplified our target DNA.  If they didn’t then we wouldn’t be able to move further in our ISHs, as there wouldn’t be the necessary hybridization.

While just running PCR might sound like an easy task, the planning, sheer number of reactions, and the multitude of people in the lab area made it take much longer than we had anticipated.  Fortunately, we were able to get it all done by dinner, after which Sarah and Ruth graciously started our electrophoresis gels.

Though I have yet to see the bands on the gel, preliminary reports (read: conversation in the hallway) from Sarah and Ruth give promising results!  After we see what happened tomorrow morning we should have a good idea of how we’re moving forward.

Until tomorrow!

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