GSM2056951: L. major H3VKO DMOG; Leishmania major; ncRNA-Seq
Small RNA was isolated from log phase Leishmania major cultures using the Qiagen miRNeasy kit, according to the manufacturer’s instructions. 5x107 cells were used for each prep. Small RNA-seq libraries were constructed by Vertis Biotechnology AG, Germany. Approximately 250ng of small RNA was poly A-tailed using poly A polymerase. The 5' triphosphate and cap structures were removed using tobacco acid pyrophosphatase. An RNA adapter was ligated to the 5' monophosphate. First strand cDNA synthesis was performed using an oligo dT adapter primer and the M-MLV reverse transcriptase. cDNAs were then PCR amplified to about 10-20 ng/uL using high fidelity DNA polymerase and purified using the Agencourt AMPure XP kit.
GEO Accession
GSM2056951