RNA-Seq of camel Camelus dromedarius neurointermediate lobe of the  pituitary
Tissue was stored in RNAlater at -80OC until extraction in a Category II  fume hood. Samples were then weighed and dissected to ensure that each was  less than the 30mg to ensure that extraction by column was maximally  efficient. Each sample was then homogenized in 1ml Trizol and the aqueous  phase separated with Chloroform. Purification relied on the RNeasyMiniKit  (Qiagen) according to manufacturers protocol. Concentration and quality of  RNA was determined using both Nano-drop and Agilent Bioanalyzer 2100 and  samples with a R.I.N>8 were used for library construction. Stranded total  RNA libraries were prepared according to IlluminaTruSeq stranded total RNA  sample preparation guide (April 2013, Rev D) for Illumina Paired-End  Multiplex Sequencing (Source Bioscience, Nottingham, UK). Ribo-Zero gold kit  was used to remove cytoplasmic and mitochondrial rRNA prior to fragmentation  and priming for first and second strand cDNA synthesis. The 3 end of  fragments were adenylated and adapters and indexes (for the multiplex  barcoding) ligated. Libraries were then subject to further quality control  prior to pools with a concentration of 8pM being loaded onto lanes of an  Illumina flowcell and sequenced using 100bp Paired End runs. Each sample  resulted in >35million reads.