I’ve spent the last few weeks trying to figure out why my
carbonic anhydrase knockdown cell line is not exhibiting social motility at 5%
CO2 during the SoMo assay. For a while, we all thought it was my technique. But
now, we all think there’s something wrong with the cell line. After I showed
the data to my P.I., he believes that I have kept these parasites in culture
for too long. Apparently, when you
passage parasites (or any other microbe for that matter), in suspension culture
for too long, they can acquire mutations or alter their gene expression levels
in a way that can disturb normal knockdown levels upon RNAi (RNA Interference)
induction with tetracycline. Wow, that was definitely a mouthful even for me! I
guess the take-away lesson is that you want always want to be working with
fresh aliquots of these parasites!
Check them out! Notice the turbidity of the media in the
flask. That’s a dense culture of parasites. Looks like it’s time to split
(dilute) them into a new flask
And also quickly, I wanted to introduce my good friend,
Hoangkim Nguyen. She’s one of the graduate students who recently published a
paper on CMF22, an axonemal protein that is required for propulsive motility in
T. brucei.
Kim is great! She has been a positive
influence over my work here in the Hill Lab this summer. I come to her for
advice on my projects, career plans, and life in general!
2 comments:
Summers are the absolute best time to accomplish research as an undergraduate. Just think, it would have taken you the whole academic year possibly to trouble shoot your problem. Thanks for the post!
DDS
15 hours a week of research during the academic year agreeably is simply not enough time to make headway in a project. Thank you for all of your wonderful support this summer, Dr. Simmons!
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