alias: E-MTAB-6281:RNA-IP-1FL_s
title: Illumina HiSeq 2000 sequencing; RNA-Seq data: RNA-IP (TAP-ZC3H30 pull down followed by RNA-Seq) and total transcriptomics (Effect of ZC3H30 gene deletion) in procyclic form of T. brucei
design: RNA-Seq data: RNA-IP (TAP-ZC3H30 pull down followed by RNA-Seq) and total transcriptomics (Effect of ZC3H30 gene deletion) in procyclic form of T. brucei
library_construction_protocol: RNA-IP protocol-Procyclic TAP-ZC3H30/SKO were grown at log phase till the density was 1.5million cells/ml and not more than 2million cells/ml. Cell culture was pulled at 1125xg for 10 minutes. It was washed with 10ml of MEM-Pros  medium (without FCS), followed by UV crosslinking at  (400mJ/square cm) while on ice, recentrifuged at 1125xg, for 10 minutes. The final cell pellet was washed with cold 1xPBS and proceeded for lysis and immunoprecipitation. T.brucei procyclic cells were grown at log phase till the density was 1.5million cells/ml and not more than 2million cells/ml. Cell pellet was washed with MEM-Pros (without FCS) and UV crosslinked 400mJ/square cm) while on ice. Log phase cell culture was heat shocked at 39 degrees for 1 hour before RNA extraction. Standard TRIFast FL protocol from PeqLab Standard Illumina library preparation protocol
identifiers.namespace: ZMBH
identifiers.id: E-MTAB-6281:RNA-IP-1FL_s