Will Wikoff, Weeks 2 and 3, Progress

After the first week at the Matsunami lab I finally completed all of my bench work training and delved into experiments with my grad student, Serene.   The more accustomed I became to the lab the more routine my schedule became as it synched up with those of my fellow lab members.   Now my average day at the lab follows this trend:

 

Arrive at the lab between 9:00-9:15am

Begin working with the lab manager, Jessica, when she arrives around 9:20am

Work on OR Library until around 12:20-12:40pm

Find time for Lunch around 12:30-1:15pm

Work with Serene from 1:30-5:15pm

The lab is empty by 5:45pm

 



The primary human OR library bin (HuOR 1)
Contains over 600 unique gene variants

In the mornings Jessica teaches me new lab skills through helping maintain the OR library.  The OR library is a group of massive bins containing thousands of different olfactory receptor genes from human and mice alike.  They require constant maintenance because, in addition to being used regularly for experiments in our lab, we share our samples with countless other labs.  For instance, last week a PI from California requested to fly over and make a copy of our entire library for their lab.  With the TE buffer the DNA is kept in constantly evaporating or the samples being used up, I am constantly transforming new cell cultures, incubating them, prepping the genes, and sequencing them– my hand eye coordination with the micropipettes has increased tenfold since week one.  The best part about all of this bench work is seeing it all come together day five when I finally get to analyze the sequencing data and see how many of my samples are added to the library.  However, the real fun begins in the afternoon when I work under Serene.

 



Working under Serene was a little slow at first.  Many of her projects are more complex than the other graduate students and as such more costly.  It took a little while before she began to trust me with sectioning +$100 mice or insert the gels for gene samples she only had five microliters of, but now I work alongside with her for all her experiments.



Sectioning a mice olfactory epithelium using a

cryostat microtome.  (Scientific deli slicer)

Running gels and making Ms. Salmon Proud

 



So what are we working on?

 

Analyzing data: looking for mutation in observed genes:
(ie. deletions, insertions, heterozygous, or homozygous)

Serene’s research pertains to innate responses associated to certain odors.  After observing that mice exhibit a flight, fight, or freeze response to fox or coyote odor she begun testing mice in order to locate the genes and mechanisms involved.  Currently we are at a stage in this research where we located three or four potential genes involved and are conducting behavioral studies on the mice.  In addition to behavioral studies afterwards we euthanize mice in order to conduct in situ hybridization to locate these receptors.  Right now I am comparing results from the gel that I ran and the gene sequencing results in order to determine whether the mice we used were mutated or wild type and if these are data points we can submit.  So far things are looking good which means we will probably conduct a behavioral study this week (the first one she has done since I arrived at the lab) and I am very excited for that.

 

Now that the first three weeks are over everything is at full throttle.  Week four is my middle week– as of Wednesday I am halfway there.