Dataset features


Application: Gene expression microarray analysis
Number of samples: 43
Release date: Jun 1 2009
Last update date: Jan 18 2013
Access: Public
Chemicals: Aminoglycosides
Dataset link Expression profiling in lymphoblastoid cell lines of family with maternally inherited non-syndromic hearing loss.

Experimental Protocol

Lymphoblastoid cell lines established from peripheral blood lymphocytes of study participants and immortalized with Epstein-Barr virus, were grown in suspension in T flasks in RPMI-1640 medium (Invitrogen, Inc., Carlsbad, CA), containing 2 mmol/L L-glutamine, 100 ug/ml streptomycin, and 10% fetal calf serum. Total RNA was extracted using Trizol reagent (Invitrogen, Inc.) according to the manufacturer?s protocol. cRNA amplification and labeling with biotin were performed using Illlumina® TotalPrep RNA amplification kit, manufactured by Ambion, Inc (Austin, TX) according to the manufacturer?s protocol using 100 ng of total RNA as input material. 1ug of labeled cRNAs were hybridized to the eight ?whole genome? Sentrix Human-6 v2 Expression BeadChips (Illumina, San Diego, CA). RNA sample from patient Unaffected 2 was duplicated on the same chip; RNA sample from Affected 3 was analyzed in triplicate: two on the same chip, and one on a different chip. All hybridization and scanning steps were performed according to the manufacturers? instructions using reagents and equipment purchased from Illumina, Inc