alias: GSM2046508
title: GSM2046508: Dark-housed Gad2-Cre 3hr light rep 3; Mus musculus; RNA-Seq
library_construction_protocol: RiboTag-purification accoriding to Spiegel et al (Cell, 2014) and Sanz et al (PNAS 2009) All samples had RIN numbers above 8.0. 5-10 ng of RNA were SPIA-amplified with the Ovation RNA Amplification System V2, yielding 5-8 μg of cDNA per sample. SPIA-amplified samples from RiboTag-IP fractions were processed in triplicate. For preparing Seq-libraries, 2 mg of each amplified cDNA were fragmented to 200-400 bp using a Covaris S2 sonicator (Acoustic wave instruments) using the following parameters: duty cycle = 10%, intensity = 5, cycles per burst = 200, time: = 60 seconds, total time = 5 minutes. After validating the fragment length of the sonicated cDNA using a 2100 BioAnalyzer, 2 mg of the fragmented cDNA were used for Seq library preparation using the PrepX DNA kit on an Apollo 324 robot (IntegenX). Quality of completed Seq-libraries was assessed using a 2100 BioAnalyzer (Agilent, Palo Alto, CA) and the completed libraries were sequenced on an Illumina HiSeq 2000 instrument per the manufacturer's standard protocols for single-end 50-bp sequencing with single-index reads. SPIA amplification of 5-10 ng ribosome bound RNA
identifiers.namespace: GEO
identifiers.id: GSM2046508