Laura Melanie BitterlichNational University of Ireland, Maynooth | NUI Maynooth · Department of Biology
Laura Melanie Bitterlich
Master of Science
About
2
Publications
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Citations
Introduction
My PhD research project focuses on the effect of the obese microenvironment on mesenchymal stromal cell immunomodulation efficacy.
Additional affiliations
October 2018 - December 2018
August 2016 - July 2017
January 2019 - July 2020
Education
September 2019 - September 2020
October 2014 - July 2018
Publications
Publications (2)
Mesenchymal stromal cell (MSC) therapy has seen increased attention as a possible option to treat a number of inflammatory conditions including COVID-19 acute respiratory distress syndrome (ARDS). As rates of obesity and metabolic disease continue to rise worldwide, increasing proportions of patients treated with MSC therapy will be living with obe...
Questions
Question (1)
EDIT: The problem turned out to not be with my isolated RNA, my synthesised cDNA, or any of the other components. Instead, I missed that the setting for the reference dye was set to "ROX" instead of "None".
I isolated RNA from cells and transcribed it into cDNA which I then used for a qPCR. In the first run, only a few wells yielded a CT, for most I received an "undefined". I re-ran the experiment with the same result, and I'm not sure what the problem could be, because:
- Both runs resulted in 10 primer-sample combinations for which I had a CT. However, those were not the same combinations both times
- All primers have at least one result with a CT
- All samples have at least one result with a CT
- The primers have worked before
- All are using the same master mix with SYBR Green (with different primers in each mix, of course) which has also worked very recently
- I'm using a template for the run parameters that has worked before
Technically, it should not be the primers, not the samples, and not the master mix. But what else could it be? I consider re-synthesising my cDNA and repeating the experiment, but I'm unsure this will fix anything.
