Hello RG community,
My task is to measure the expression of several microRNAs from total RNA isolated from human body sera. I have a number of TIPS patient (suffering from liver cirrhosis) and healthy control RNA samples. I was told that the patient RNA was isolated from sera of the liver vein but the control samples were isolated from sera of peripheral blood (arm vein) from healthy humans.
Basically, I perform microRNA Revers Transcription using a miR-specific primer and a stem-loop primer. Later, a microRNA qPCR is carried out and relative quantification is achieved by comparing the ct of the miRNA of interest to the spike in control´s ct value.
After all, my question is if the results are valueable because we compare the expression of circulating microRNAs initially obtained from to different body compartments ?
My previous field of study was Microbiology so I do not have enough experience which is why I need to place this question.
I´d be extremely grateful for any professional opinion and guidance to my problem.
Thanks to you in advance !!
Hi
U will use qrtpcr for fold changes values.for ur control and test.u will use ddct method to get rq values that need to be seen for statistics to confirm significance.if mirna get differentially regulated then u will search potential targets here in ur case cirrhosis associated gene.u may have to do mirbase gene search and put it into gene ontology software to predict the go then u will sort go of ur relevance say cirrhosis then will get possible role of these mirs in ur patiant
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