Transcriptomics_in_antigenically_selected_lines_of_Plasmodium_falciparum_-sc-2012-05-23T13:02:48Z-2224
Transcriptomics_in_antigenically_selected_lines_of_Plasmodium_falciparum_
FRT-Seq is an alternative approach to transcriptome sequencing in which the reverse transcription reaction takes place on the flowcell. No amplification is performed during the library preparation, so PCR biases and duplicates are avoided, and because the template is poly(A)+ RNA rather than cDNA, the resulting sequences are necessarily strand-specific. Here we have used FRT-Seq to study gene expression in populations of Plasmodium falciparum parasite clones which were selected for homogenous expression of single var genes. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/ Protocols: Parasites were cultured according to the method of Trager and Jensen (Human Malaria Parasites in Continuous Culture. Science. 1976 Aug 20;193(4254):673–5). Cultures were synchronized by sorbitol lysis (to obtain enrichment of ring stage parasites). The IT clone and its subclones were used for phenotypic selection experiments. The A4 subclone was positively and negatively selected for expression of the BC6 antibody. RNA extraction was performed with Trizol (Invitrogen) according to manufacturer instructions. FRT-Seq was performed as described by Mamanova et al.  (FRT-seq: amplification-free, strand-specific transcriptome sequencing. Nature Methods. 2010 Jan 17;7(2):130–2).
FRT-Seq is being carried out to study transcriptomics in antigenically selected lines of Plasmodium falciparum.