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Global Gene Expression Profiles of Skeletal Muscle in Males with Type 2 Diabetes.

ABSTRACT: Gene expression profiles of biopsy samples of skeletal muscle of three male patients of type 2 diabetes and three non-diabetic male patients were generated using Illumina HumanHT-12 v3 Expression BeadChip arrays. The primary indications of surgery were non-infective and non-malignant conditions, namely, cholelethiasis, hernia and trauma. Three biological replicates with four technical replicates each were used to generate expression profiles. Total cellular RNA from biopsy samples was isolated using mirVana™ miRNA Isolation Kit (Ambion, Austin, TX). The quantity and the quality of RNA samples were determined using Nanodrop-1000 (Thermo Fischer Scientific, Wilmington, USA) and Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA), in that order. RNA samples with a RIN (RNA Integrity Number) value between 5-8 were used for expression profiling. Starting with 500 ng of total RNA, Illumina TotalPrepTM RNA Amplification Kit was used for preparing first and second strand cDNA, purification of cDNA, in vitro transcription to synthesize biotin labeled cRNA, and purification of the labeled cRNA, in that sequence. The quantitation of cRNA was performed using Nanodrop-1000. Illumina HumanHT-12 v3 Expression BeadChip arrays were hybridized with 750 ng of labeled cRNA samples. Hybridization and washing were perforemd according to the manufacturer's protocol. The arrays were scanned and read using Illumina iScan System, and the data extraction, average normalization and downstream analysis performed using Illumina GenomeStudio V2010.1.

ORGANISM(S): Homo sapiens

SUBMITTER: Abhay Sharma

PROVIDER: E-GEOD-29221 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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