GSM5859174: Shh KO rep5; Mus musculus; RNA-Seq
pSHFs were microdissected, flash frozen on dry ice, and RNA was harvested using Trizol reagent and RNeasy Mini RNA Isolation Kit (Qiagen 74104). TruSeq RNA Sample prep kit v2 (Illumina RS-122-2001) was used with 1 ug of total RNA for the construction of sequencing libraries. Libraries were quantitated on an Agilent Bio-Analyzer and pooled in equimolar amounts. Pooled libraries were sequenced on the HiSeq2500 in Rapid Run Mode following the manufacturer's protocols to generate stranded single-end reads of 50bp.