Transcriptome Analysis and Discovery of Genes Involved in Immune Path... - SRA

SRX381027: Transcriptome Analysis and Discovery of Genes Involved in Immune Pathways of Microbial Challenged musca domestica
1 ILLUMINA (Illumina HiSeq 2000) run: 52.3M spots, 10.5G bases, 6.6Gb downloads

Design: A total amount of 3ug RNA per sample was used as input material for the RNA sample preparations. All four samples had RIN(RNA Integrity number) values above 8. Sequencing libraries were generated using NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB, Ispawich, USA) following manufacturer’s recommendations and four index codes were added to attribute sequences to each sample. Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads (Life technologies, CA, USA). Fragmentation was carried out using divalent cations under elevated temperature in NEB proprietary fragmentation buffer. First strand cDNA was synthesized using random oligonucleotides and M-MuLV Reverse Transcriptase (RNase H-). Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. Remaining overhangs were converted into blunt ends via exonuclease/polymerase activities and enzymes were removed. After adenylation of 3’ ends of DNA fragments, NEBNext adaptor oligonucleotides were ligated to cDNA fragments. In order to select cDNA fragments of preferentially 200 bp in length the library fragments were purified with AMPure XP beads system (Beckman Coulter, Beverly, USA). DNA fragments with ligated adaptor molecules on both ends were selectively enriched using NEB Universal PCR Primer and Index primer in a 10 cycles PCR reaction. Products were purified (AMPure XP beads system) and quantified using the Agilent high sensitivity DNA assay on the Agilent Bioanalyzer 2100 system.

Submitted by: INSTITUTE OF ANIMAL HUSBANDRY AND VETERINARY SCIENCE,HENAN ACADEMY OF AGRICULTURAL SCIENCES (INSTITUTE OF ANIMAL HUSBANDRY AND VETERINARY SCIEN)

Study: Musca domestica Transcriptome or Gene expression

PRJNA227268 • SRP033226 • All experiments • All runs

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Transcriptome Analysis and Discovery of Genes Involved in Immune Pathways of bacteria Challenged musca domestica

Sample: Transcriptome sequencing of Musca domestica larvae

SAMN02401116 • SRS505438 • All experiments • All runs

Organism: Musca domestica

Library:

Instrument: Illumina HiSeq 2000

Strategy: RNA-Seq

Source: TRANSCRIPTOMIC

Selection: RT-PCR