title: GSM2175278: Trypanosoma brucei brucei Lister 427, rep1; Trypanosoma brucei; OTHER
library_construction_protocol: gDNA extraction then PCR amplificaiton of the RNAi inserts An RNAi library population of 2x107 cells (~5 fold genome coverage) was selected in the presence of 100µM 8-pCPT-2'-O-Me-cAMP or 10µM 8-pCPT-2'-O-Me-5'-AMP22 in several replicate flasks either induced, or not, to activate RNAi.   Three cAMP-selected and five AMP-selected populations outgrew and were subject to DNA isolation and RNAi fragment insert amplification. Amplicon populations from the cAMP and AMP selections were analysed by ion torrent sequencing.   PCR amplicon primers (Forward primer: 5' TAGCCCCTCGAGGGCCAGT 3', Reverse primer: 5' GGAATTCGATATCAAGCTTGGC 3') were used to generate PCR fragments, which ranged in size up to about 2kb. These were subsequently fragmented to 250-200nt sized fragments before barcoding and sequencing.
attributes.tag: GEO Accession
attributes.value: GSM2175278