GSM2670889: Adult_CD235a_RNA_m_1; Homo sapiens; RNA-Seq
Total RNA was extracted using Qiagen RNeasy spin columns and subsequent poly(A)+ selection was performed using the NEB NEXT kit by following the manufacturers instructions. Poly(A)+ selected RNA was fragmented to ~200nt by a 3 min. incubation at 95 °C in FAST-AP buffer. Samples were quickly transferred to ice and subjected to joint Alkaline Phosphatase (FAST-AP, Thermo Scientific) and T4 Polynucleotide Kinase (PNK, NEB) treatment for end-repair. Following incubation at 37 °C for 30 min, reactions were cleaned with MyOne SILANE magnetic beads, washed twice with 70% ethanol and eluted in 6 μl H2O. Library preparation was carried out using an Illumina TruSeq Kit, as per manufacturer’s recommendation, and samples were sequenced on the Illumina HiSeq 2500
GEO Accession
GSM2670889