Carausius morosus

Transcriptome and Neuropeptidome Analysis of Carausius morosus

RNA isolation, cDNA preparation and Illumina Next Generation Sequencing (per manufacturer’s protocol, Illumina, San Diego, CA) were carried out by BGI (New Territories, Hong Kong). Each library was sequenced for 100 bp paired-ends, which were stored as FASTQ files. The resulting RAW reads were filtered, by removing adapter sequences, contamination and low-quality reads (clean reads: 49784152, clean bases: 4978415200, Q20%: read1: 99.11; read2:97.92).The overall size of each cleaned dataset was between 6 and 7 GB. (detailed information can be obtained from section 'material and methods')

{'age': 'adult stage', 'sex': 'female', 'host': '7022', 'tissue': 'central nervous system (CNS)', 'isolate': 'adult female', 'geo_loc_name': 'Germany: colony maintained at the University of Cologne, Cologne', 'biosamplemodel': 'Invertebrate', 'collection_date': '2016-05-03'}

RNA-seq of adult Carausius morosus female

quaility control, poly-A selection, library preparation, Rapid Run sequencing

{'sex': 'female', 'tissue': 'Total body', 'ecotype': 'Indian', 'dev_stage': 'Adult', 'phenotype': 'wild type', 'sample_type': 'tissue sample', 'collected_by': 'Burcin Duan Sahbaz', 'biosamplemodel': 'Model organism or animal', 'breeding_method': 'ad libitum', 'collection_date': '2009-08'}

Neuromodulation can be simple: myoinhibitory peptide, contained in dedicated regulatory pathways, is the only neurally-mediated peptide modulator of stick insect leg muscle

Legs were dissected as described above. RNA was extracted from the ExtTi and FlxTi muscles of a single middle leg using TRIzol (Thermo Fisher Scientific) following manufacturer recommendations. RNA quality was estimated using RNA concentration (ng/μL) and RNA integrity number as implemented in Agilent 2100 Bioanalyzer. Libraries were prepared using 1 μg total RNA with the Illumina® TruSeq® stranded RNA sample preparation kit. PolyA containing mRNA were purified using oligo-dT attached magnetic beads and then fragmented and primed for cDNA synthesis. Fragmented RNA was reverse transcribed using random hexamers to produce first strand cDNA; second strand cDNA was synthesized with DNA polymerase I and RNase H. After end repair and A-tailing, multiple indexing adapters for hybridization onto a flow cell were ligated. Products were enriched with PCR and purified to create the final cDNA library. Sequencing was done on an Illumina HiSeq 4000 sequencer with 75 paired ends at Cologne Center for Genomics (CCG, Cologne, Germany).

{'sex': 'female', 'tissue': 'muscle', 'dev_stage': 'adult', 'collected_by': 'Sander Liessem', 'biosamplemodel': 'Model organism or animal', 'birth_location': 'Cologne', 'biomaterial_provider': 'Bueschges Lab, University of Cologne'}