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SRX19062607: GSM6940588: WT biol rep1; Plasmodium yoelii yoelii 17XNL; RNA-Seq
1 ILLUMINA (Illumina NovaSeq 6000) run: 20.7M spots, 6.2G bases, 1.9Gb downloads

External Id: GSM6940588_r1
Submitted by: Xiamen University
Study: RNA sequencing of Rbpm1 knockout in Plasmodium yoelii
show Abstracthide Abstract
RBPm1, an RNA-binding protein, plays a crucial role in regulating axoneme assembly during male gametogenesis of Plasmodium yoelii. To elucidate the molecular mechanisms that underlie this process, we conducted RNA-seq analysis to investigate changes in the male transcriptome resulting from the loss of RBPm1. Overall design: To obtain highly pure male gametocytes, we employed a DFsc7 reporter strain that selectively exhibited GFP expression within these cells. We created a mutant strain by deleting RBPm1 from the DFsc7 strain. Male gametocytes from both the DFsc7 and mutant strains were sorted using fluorescence-activated cell sorting (FACS), relying on GFP fluorescence intensity as a criterion, and subsequently subjected to RNA-seq analysis. Addition: Through Principal component analysis (PCA), it was observed that the RNA-seq data from the initial sample, WT biol rep1, exhibited poor reproducibility. Subsequently, we re-sorted and obtained three additional WT samples (WT_biol_rep4, WT_biol_rep5, and WT_biol_rep6) for RNA-seq, resulting in improved data reproducibility.
Sample: WT biol rep1
SAMN32782854 • SRS16478289 • All experiments • All runs
Library:
Name: GSM6940588
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Total RNA from 2×10^7sex-sorted gametocytes was extracted using TRIzol. RNA integrity was checked with the Agilent 2100 Bioanalyzer. mRNA was then enriched by Oligo(dT) beads. The enriched mRNA was fragmented into short fragments using fragmentation buffer and reverse transcripted into cDNA with random primers. Second-strand cDNA were synthesized by DNA polymerase I, RNase H, dNTP and buffer. The cDNA fragments were purified with the QIAQuick PCR Purification Kit, end-repaired, A tailed, and ligated to Illumina sequencing adapters. The ligation products were size-selected by agarose gel electrophoresis, PCR amplified, and sequenced using Illumina NovaSeq 6000 by Genedenovo Biotechnology Co., Ltd (Guangzhou, China).
Runs: 1 run, 20.7M spots, 6.2G bases, 1.9Gb
Run# of Spots# of BasesSizePublished
SRR2311125320,652,9256.2G1.9Gb2024-04-18

ID:
26234071

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