NextSeq 500 paired end sequencing; Cardiac miRNA expression changes induced by prediabetes
Cardiac miRNA expression changes induced by prediabetes
At week 21 of the diet, animals were anesthetized with pentobarbital (60 mg/kg, i.p.), then hearts were excised, shortly perfused with oxygenated Krebs-Henseleit buffer in Langendorff mode and stored at -80°C in RNAlater. RNAlater-preserved rat left ventricle samples (n=6/group) were thawed on ice, then ca. 30 mg of left ventricular tissue were placed in a 1.5 ml Eppendorf LoBind tube containing glass beads (1.7-2.1 mm diameter, Carl Roth, Karlsruhe, Germany) and 500 µl of VRX buffer (Viogene Biotek, New Taipei City, Taiwan, R.O.C). The Eppendorf tube was firmly attached to a SILAMAT S5 vibrator (Ivoclar Vivadent, Schaan, Liechtenstein) in order to disrupt and homogenize the tissue for 15s. Total RNA was extracted using Viogene miTotal RNA Extraction Miniprep System (Viogene Biotek, New Taipei City, Taiwan, R.O.C) according to the manufacturer's protocol. For small RNA library construction, NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England Biolabs, Ipswich, MA, USA) was applied according to the manufacturer's protocol. The quality and quantity of the library QC was performed by using High Sensitivity DNA1000 ScreenTape system with 2200 Tapestation (Agilent Technologies, Santa Clara, CA, USA) and dsDNA HS Assay Kit with Qubit 3.0 Fluorometer (Thermo Fisher Scientific, Waltham, MA, USA), respectively.
Experimental Factor: disease
normal
Experimental Factor: compound
none