title: GSM1470698: PLK_pH7.2; Toxoplasma gondii; RNA-Seq
library_construction_protocol: RNA was extracted using Invitrogen TRIzol® Reagent (cat#15596018), followed by genomic DNA removal and cleaning using Qiagen RNase-Free DNase Set kit (cat#79254) and Qiagen Mini RNeasy™ kit (cat#74104). Agilent 2100 Bioanalyzer was used to assess the integrity of the RNA samples. Only RNA samples having RNA Integrity Number between 9-10 were used. Ambion® MicroPoly(A)Purist™ Kit (cat#AM1919) was used for enrichment of transcripts.  The SOLiD® Total RNA-Seq Kit (cat#4445374) was used to construct template cDNA for RNA-Seq following the protocol recommended by Applied Biosystems. Briefly, mRNA was fragmented using chemical hydrolysis followed by ligation with strand specific adapters and reverse transcript to generate cDNA.  The cDNA fragments, 150 to 250 bp in size, were subsequently isolated by electrophoresis in 6% Urea-TBE acrylamide gel. The isolated cDNA was amplified through 15 amplification cycles to produce the required number of templates for the SOLiD™ EZ Bead™ system, which was used to generate template bead library for the ligation base sequencing by the SOLiD™4 instrument.