title: GSM1177000: Purine-Depleted sample; Leishmania donovani; RNA-Seq
library_construction_protocol: 1 X 10^8 cells at early log phase were removed from each condition, harvested by centrifugation and washed twice in D-PBS to remove all media components.  Total RNA from the purine-replete and purine-deplete cells was isolated using an RNeasy kit (Qiagen Inc., Valencia, CA) according to the manufacturer's protocol. Next Generation Sequencing (NGS) libraries were constructed by priming ~1 µg of total RNA from either the purine-starved or purine-replete sample with a random hexamer primer (TCCGATCTCTNNNNNNN) for first strand cDNA synthesis and an oligonucleotide primer (TCAGTTTCTGTA) derived from the Leishmania splice leader (SL) sequence for second-strand cDNA synthesis.  The resultant cDNA was amplified by PCR for 12-15 cycles using the oligonucleotides that overlapped with the initial primers and containing adapters for subsequent cluster generation and sequencing using Solexa sequencing by synthesis (SBS) technology.