GSM1366347: dsRNA in DICER kd cells; Homo sapiens; RNA-Seq
total RNA was prepared using Trizol reagent, digested using T1 nuclease and resulting dsRNA was sequenced 100ng of sample was phosphatase and PNK treated, as in the Illumina mRNA Directional protocol. All clean ups (post-PNK, post-PCR) were done using Ampure XP beads. Following steps were carried out using the NEBNext Small RNA Library Prep Kit but with custom indexes designed by the WTCHG facility (Reference: Improved workflows for high throughput library preparation using the transposome-based nextera system <http://www.biomedcentral.com/1472-6750/13/104> Sarah Lamble, Elizabeth Batty, Moustafa Attar, David Buck, Rory Bowden, Gerton Lunter, Derrick Crook, Bassam El-Fahmawi, Paolo Piazza BMC Biotechnology 2013, 13:104, 20 November 2013). The concentration of each library was determined by realtime using Agilent qPCR Library and a MX3005P instrument (Agilent). Sequencing was performed on an Illumina HiSeq2000, using 50bp paired end reads.
GEO Accession
GSM1366347