Identification of cardiomyocyte-derived microvesicles content.. Mus musculus

Investigation of content of microvesicles exocytosed by cardiomyocytes. The aims of the study were to identify DNA and RNA content in microvesicles from cardiomyocytes. DNA and RNA were purified from microveicles from cultured cardiomyocytes. Illumina microarrays were used to detect DNA and mRNA molecules and identify them. Overall design: Gene expression microarray were used to detect 50 nucleotide long illumina probe sequences in the sequences of the extracted microvesicular DNA. Terminal deoxynucleotidyl transferase were used to add a poly_T tail to DNA from microvesicles to construct cDNA, possible to label as cRNA and use on the chip to identify part of the sequence of the original DNA. The microvesicular mRNA content was compared to cardiomyocyte mRNA content (the source of the microvesicles and therefore also the source of microvesicle mRNA). Two replicates of mRNA from cardiomyocytes and two replicates of mRNA from microvesicles and one sample of DNA from microvesicles were analyzed.