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.