title: GSM1303493: hom_RNF17_5RACE_small and large500bp; Mus musculus; RNA-Seq
library_construction_protocol: total RNA was extracted from manually dissected testes with Trizol reagent (Invitrogen) 5'RACE libraries were prepared according  (Karginov et al. 2010, Mol Cell 38: 781-788).  Poly(A)+ mRNAs were isolated from 100 g of total RNA using the Invitrogen Dynabeads mRNA Direct kit. The RNA was ligated to a 5 linker with T4 RNA ligase (Ambion). Reverse transcription reactions were carried out by handom hexamer priming SBS8-N6 primer and SuperScript III (Invitrogen). Template RNA was degraded by addition of 1 l RNase H (Invitrogen). PCR reactions on the resulting cDNAs were carried out using KOD Hot Start DNA polymerase (Novagen) with PE-P5-SBS3 and PE-P7-SBS8 primers. Products were run on a 2% low-melt agarose gel, and the 150500 bp (sometimes 1501000bp) range was purified.
attributes.tag: GEO Accession
attributes.value: GSM1303493