GSM7987501: NC3; Homo sapiens; RNA-Seq
The stem cells in a high-glucose environment, treatment with high levels of glucose for 48 hours, hUCMSCs were subjected to high-throughput RNA sequencing to obtain the transcription profile of these cells under high-glucose conditions.  Sample preparation and microarray hybridization are carried out according to Arraystar's standard protocol. Simply put, the total RNA isolated from each sample is amplified and transcribed into fluorescent RNA using random primers, and the array is scanned with an Axon GenePix microarray scanner. Then the scanned images were imported into GenePix and data extraction was carried out. LncRNAs with significant differentially expressed expression were screened by fold change > 2 or < -2 and P value < 0.05 RNA-seq libraries were constructed by NewCore Biotech (Shanghai, China). Firstly, RNA was isolated with TRIZOL reagent (Invitrogen, Grand Island, NY), total RNA was purified by the Qiagen RNeasy mini kit (Qiagen, Valencia, CA). Purified RNA was analyzed on an ND-8000 spectropho tometer (Nanodrop Technologies, Wilmington, DE). High quality total RNA was used as the starting material. Briefly, total RNA was used for removing the rRNAs with NEBNext rRNA Depletion Kit (New England Biolabs, Inc., Massachusetts, USA) following the manufacturer's instructions. RNA libraries were constructed by using NEBNext® Ultra™ II Directional RNA Library Prep Kit (New England Biolabs, Inc., Massachusetts, USA) according to the manufacturer's instructions. Libraries were controlled for quality and quantified using the BioAnalyzer 2100 system (Agilent Technologies,Inc., USA). Library sequencing was performed on an Illumina NovaSeq 6000 instrument with 151 bp paired end reads.