GSM1481532: mother; Homo sapiens; RNA-Seq
RNA was extracted from the peripheral blood mononuclear cell (PBMC) layer derived by Ficoll-paque extraction from 20 mL of whole blood. In the case of the proband, blood was drawn immediately prior to transfusion to enrich for endogenous nucleated erythroid cells and reticulocytes. Beginning with 20 µg total RNA, the Ambion GLOBINclear™ kit (#AM1980, Life Technologies) was used to remove human alpha-globin and beta-globin mRNA. Ambion Dynabeads® mRNA DIRECT™ Micro Purification Kit (#61021, Life Technologies) was used to select the polyA+ (mRNA) fraction. ERCC control RNA from the Ambion ERCC RNA Spike-In Mix Kit (#4456740, Life Technologies) was added to estimate the amplification efficiency during library construction. Barcoded RNA-seq libraries were prepared using the Ion Total RNA-Seq Kit v2 (#4475936, Life Technologies) along with the Ion Xpress™ RNA-Seq Barcode 1-16 Kit (#4475485, Life Technologies) for sequencing on the Ion Proton instrument (Life Technologies). Three barcoded libraries were sequenced on a single P1 chip such that approximately 20 million mapped reads were obtained for each sample.
GEO Accession
GSM1481532