![]() ![]() ![]() The 3'-end non-bias proportional RNA amplification was carried out using the TransPlex™ Whole Transcriptome Amplification kit (Rubicon Genomics, Ann Arbor, MI).Įach sample was incubated in the dark at 37✬ for 1 h with 120 U Terminal Deoxynucleotidyl Transferase Recombinant Enzyme and Terminal Transferase Buffer (Promega, Madison, WI), and 2.4 nmol Cy-3 dUTP (Amersham Biosciences, Little Chalfont Buckinghamshire, England). Cells were laser-capture microdissected with the Arcturus PixCell II system and collected.Ĭellular RNA was extracted using the PicoPure RNA isolation kit (Arcturus). ![]() Ventricular myocytes were identified in serial 7-8-µm thick cryostat sections using the RNA-preserving quick (10 s) staining by Eosin Y (Sigma-Aldrich, St. Samples were washed in 4☌ saline, immersed in Tissue-Tek OCT compound (EMS, Hatfield, Pennsylvania), flash frozen in liquid nitrogen and stored at -80˚C until use. LLeft ventricle tissue samples were obtained from material resected from the explanted heart of an ischemic cardiomyopathy patient undergoing cardiac transplantation according to routine clinical protocols. Tissue status: freshly resected tissue sample GEO help: Mouse over screen elements for information.
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