design: 1 X 10^8 cells were 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 g 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.