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Two RNA-seq studies, one of coding and the other of coding and non-coding RNA for Identification of meningococcal factors required for colonization of epithelial and endothelial cells by high throughput screening

ABSTRACT: RNA-Seq analyses were performed in N. meningitidis strain Z5463 by deep sequencing using Ion Torrent technology (Thermo Fischer Scientific). For whole transcriptome libraries (mRNA+sRNA), cDNA libraries were prepared using the Ion Total RNA-Seq Kit v2 (Life Technologies) including a prior step of ribodepletion using specific probes adapted to N. meningitidis (Low Input Ribo Minus Eukaryote System v2) and size fractionation of RNA prior to cDNA synthesis with RNase III was performed. Briefly, 500 ng of total RNA were ribodepleted and then fragmented. After fragmentation, mean size of fragment was around 120 nt. Coding_and_NonCoding_R1, Coding_and_NonCoding_R2, and Coding_and_NonCoding_R3 represent three technical replicates. For sRNA libraries (sRNA), three cDNA libraries were prepared using the Ion Total RNA-Seq Kit v2 for Small RNA Libraries (Life Technologies) including a prior step of enrichment in small RNA fraction. Equal amounts of total RNA were used for the generation of all cDNA libraries. The three whole transcriptome libraries libraries (mRNA+sRNA) were then sequenced on an Ion ProtonTMSystem using the Ion PI Template and Sequencing OT 200 kit v3 and the 3 sRNA libraries were sequenced on an Ion PGM with the Hi-Q Template and Sequencing Kit using a 316 v2 array. NonCoding_R1, NonCoding_R2, and NonCoding_R3 represent three technical replicates.

INSTRUMENT(S): Ion Torrent Proton

ORGANISM(S): Neisseria meningitidis

SUBMITTER: Thomas Nussbaumer

PROVIDER: E-MTAB-4768 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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