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SRX2239162: RNA-Seq of D. albomicans: male 3rd instar larva
1 ILLUMINA (Illumina HiSeq 2000) run: 13.1M spots, 2.4G bases, 1.5Gb downloads

Design: We extracted total RNA from dissected tissues (Qiagen) and prepared RNA-seq libraries following the standard Illumina protocol. Briefly, we used Dynal oligo(dT) beads (Invitrogen) to isolate poly(A) mRNA from the total RNA samples. We then fragmented the mRNA by using the RNA fragmentation kit from Ambion, followed by first- and second-strand cDNA synthesis using random hexamer primers (Invitrogen). We complemented the cDNA synthesis by an end repair reaction using T4 DNA polymerase and Klenow DNA polymerase for 30 min at 20ĄC. We then added a single A base to the cDNA molecules by using 3'-to-5' exo-nuclease and ligated the Illumina adapter. The fragments were subjected to size selection on a 2% gel and purification (Qiagen). We finally amplified the cDNA fragments by PCR reaction and examined the libraries by Bioanalyzer (Aglient).Ę
Submitted by: UC Berkeley
Study: Alternative Splicing Within and Between Drosophila Species, Sexes, Tissues, and Developmental Stages
show Abstracthide Abstract
Alternative pre-mRNA splicing (“AS”) greatly expands proteome diversity, but little is known about the evolutionary landscape of AS in Drosophila and how it differs between embryonic and adult stages or males and females. Here we study the transcriptomes from several tissues and developmental stages in males and females from four species across the Drosophila genus. We find that 20-37% of multi-exon genes are alternatively spliced. While males generally express a larger number of genes, AS is more prevalent in females, suggesting that the sexes adopt different expression strategies for their specialized function. While the number of total genes expressed increases during early embryonic development, the proportion of expressed genes that are alternatively spliced is highest in the very early embryo, before the onset of zygotic transcription. This indicates that females deposit a diversity of isoforms into the egg, consistent with abundant AS found in ovary. Cluster analysis by gene expression (“GE”) levels show mostly stage-specific clustering in embryonic samples, and tissue-specific clustering in adult tissues. Clustering embryonic stages and adult tissues based on AS profiles results in stronger species-specific clustering, suggesting that diversification of splicing contributes to lineage-specific evolution in Drosophila. Most sex-biased AS found in flies is due to AS in gonads, with little sex-specific splicing in somatic tissues.
Sample:
SAMN05897922 • SRS1738883 • All experiments • All runs
Library:
Name: alb_m_larva
Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Runs: 1 run, 13.1M spots, 2.4G bases, 1.5Gb
Run# of Spots# of BasesSizePublished
SRR441617413,148,6712.4G1.5Gb2016-11-15

ID:
3283833

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