title: GSM2881429: 1_2i_empty; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted using an RNeasy mini kit (Qiagen) and treated with DNase (Ambion AM1907). The RNA was quality checked on a 2200 Tapestation using the RNA ScreenTape assay (Agilent Technologies, Wokingham, UK) and the RNA concentration measured using a Qubit RNA High Sensitivity kit (LifeTechnologies, Paisley, UK). For ChIP-seq, After reverse-crosslinking, DNA was purified with a minElute PCR purification kit. PolyA-tailed mRNA was separated and libraries were prepared using KAPA's Stranded mRNASeq kit (Roche Diagnostics, Burgess Hill, UK) according to the manufacturer's instructions using an input of 500 ng, a fragmentation incubation time of 6 minutes at 94°C, and 12 cycles of PCR for library amplification. Samples were sequenced on Illumina's NextSeq500 (Illumina Cambridge, Chesterford, UK) using a high output 300 cycle paired-end run in order to map repetitive regions more accurately. Twenty-four libraries were multiplexed in the same run to achieve up to 16M reads/sample. Libraries were pooled in equimolar quantities, calculated from concentrations measured using the Qubit dsDNA High Sensitivity assay kit (Life Technologies, Paisley, UK) and fragment analysis using the D1000 High Sensitivity kit on the 2200 Tapestation (Agilent Technologies, Wokingham, UK).  For ChIP-seq, we used the NEBNext DNA Ultra II library prep kit from New England Biolabs. 1ng was used and 10 cycles of amplification were performed. Single 86bp read sequencing was performed with >100 million reads per sample. Stranded mRNA-seq. For ChIPseq: single 86 bp read sequencing
attributes.tag: GEO Accession
attributes.value: GSM2881429