Cardiac RNA Extraction and cDNA Synthesis

RNA was extracted using Qiagen RNeasy kits (Qiagen, Hilden, Germany) from the viable myocardium of the remote noninfarct region of the left ventricle (30 mg) from MI rats. This region of the myocardium was chosen because it receives even exposure to uremic toxins concentrations observed in the circulation. A similar region of the heart was obtained for sham rats (Lekawanvijit et al. 2013 (link)). Then cDNA synthesis was performed using microRNA-specific primers (Applied Biosystems, Foster City, CA), by method provided by the manufacturer. Standard cDNA synthesis protocol was followed to obtain cDNA for gene assays (Nankervis et al. 2013 (link)).

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Rana I., Kompa A.R., Skommer J., Wang B.H., Lekawanvijit S., Kelly D.J., Krum H, & Charchar F.J. (2015). Contribution of microRNA to pathological fibrosis in cardio-renal syndrome: impact of uremic toxins. Physiological Reports, 3(4), e12371.

Publication 2015

RNeasy kits microRNA-specific primers

Assays Cdna Gene Heart Left ventricle Microrna Myocardium Primers Rats Synthesis Uremic toxins

Corresponding Organization : Federation University

Other organizations : Monash University, St Vincent's Hospital, University of Melbourne

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Variable analysis

independent variables

Myocardial infarction (MI) in rats

dependent variables

Gene expression in the viable myocardium of the remote non-infarct region of the left ventricle

control variables

Sham rats (without myocardial infarction)

controls

Positive control: None specified
Negative control: Sham rats (without myocardial infarction)

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