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SRX3530806: GSM2912656: H37Ra_4h; Homo sapiens; RNA-Seq
1 ILLUMINA (Illumina HiSeq 3000) run: 23.3M spots, 1.2G bases, 433.8Mb downloads

Submitted by: NCBI (GEO)
Study: RNA-seq identifies a distinct response in macrophages infected with Mycobacterium tuberculosis for a serial time points.
show Abstracthide Abstract
Purpose: The goals of this study are to obtain the NGS-derived transcriptome profiling (RNA-seq) for THP-1 macrophages response to Mycobacterium tuberculosis (H37Rv and H37Ra) Methods: mRNA and long noncoding RNA profiles of THP-1 macrophages infected with H37Rv and H37Ra for 1, 4, 12, 24, 48 hours were generated by deep sequencing, using Illumina Hiseq3000. The sequence reads that passed quality filters were first mapped to the latest UCSC transcript set using Bowtie2 (version 2.1.0). Then the gene expression level was estimated using RSEM (RNA-Seq by Expectation Maximization, v1.2.15), for lncRNA analysis, reads were mapped to lncRNA transcript set from LNCipedia.org. The sequence reads were normalized with TMM (trimmed mean of M-values) to identify differentially expressed genes (DEGs) using the edgeR package edgeR. qRT–PCR validation was performed using SYBR Green assays. Results: Using an optimized data analysis workflow, we mapped about 20 million sequence reads per sample to the human genome (GRCh38/hg38) and identified 25,343 mRNA and 47877 long non-coding RNA transcripts. Conclusions: Our study represents the detailed analysis of transcriptomes for THP-1 macrophages response to H37Rv and H37Ra, generated by RNA-seq technology. Overall design: NGS-derived transcriptome profiling (RNA-seq) for THP-1 macrophages response to Mycobacterium tuberculosis H37Rv and H37Ra.
Sample: H37Ra_4h
SAMN08290480 • SRS2806334 • All experiments • All runs
Organism: Homo sapiens
Library:
Instrument: Illumina HiSeq 3000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: The total RNA were isolated with RNeasy mini kit (Qiagen, Germany) according to the manufacturer's instructions Both rRNA and mitochondrial RNA was removed with Ribo-Zero Gold Removal Magnetic Kit for human/mouse/rat (Epicentre, now Illumina) according to the manufacturer's instructions. The libraries were prepared with KAPA RNA-Seq Kit for Illumina (Kapa biosystems, USA) following the manufacturer's procedure. The RNA was fragmented to the desired size (about 300 bp) by heating in the presence of Mg++. The 1st strand cDNA was synthesized with random primers. The 2nd strand cDNA synthesis converted cDNA:RNA hybrid to double strand cDNA (dscDNA), while marking the 2nd strand with dUTP. Then dAMP was added to 3'-end of dscDNA fragments. Following by ligation of 3'-dTMP adapters (10 nM) to 3'-dAMP library fragments, and PCR amplification (14 cycles) of the adapter-ligated library DNA. The dUTP-marked strand was not amplified. Library fragment size distribution was confirmed by electrophoresis and library concentration was determined with Qubit 2.0 Fluorometric Quantitation (Thermo Fisher Scientific, USA). These libraries were sequenced using the Illumina Hiseq 3000 Sequencer (50 cycles, single read lane).
Experiment attributes:
GEO Accession: GSM2912656
Links:
Runs: 1 run, 23.3M spots, 1.2G bases, 433.8Mb
Run# of Spots# of BasesSizePublished
SRR643943123,326,7281.2G433.8Mb2021-01-04

ID:
4906086

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