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SRX4665295: GSM3381512: SD1-14; Homo sapiens; RNA-Seq
1 ILLUMINA (Illumina HiSeq 2500) run: 33.3M spots, 6.7G bases, 3.3Gb downloads

Submitted by: NCBI (GEO)
Study: En bloc and segmental deletions of human XIST reveal X chromosome inactivation-involving RNA elements
show Abstracthide Abstract
RNA-seqs followed by whole and segmental deletions of XIST genes in human K562 cells. The XIST RNA is a non-coding RNA that induces X chromosome inactivation (XCI). Unlike the mouse Xist RNA, how the human XIST RNA controls XCI in female cells is less well characterized, and the XCI-involving RNA elements remain unclear. To systematically decipher the XCI-involving elements of XIST RNA, ten smaller XIST segments, including repeats A, D, and E; human-specific repeat elements; the promoter; and non-repetitive exons, as well as the entire XIST gene, were homozygously deleted using the Cas9 nuclease and paired guide RNAs at high efficiencies, followed by high-throughput RNA sequencing and fluorescence in-situ hybridization experiments on XIST RNA. Overall design: There are mock transfected cells as control and 24 clones containing whole and segmental deletion of XIST.
Sample: SD1-14
SAMN10025276 • SRS3760323 • All experiments • All runs
Organism: Homo sapiens
Library:
Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Total RNA was extracted from cells using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's instructions. Libraries were prepared according to illumina's instruction accompanying the Truseq stranded mRNA prep kit. Briefly, The polyA containing mRNA molecules were purified using poly-T oligo attached magnetic beads and then fragmented into small pieces. The cleaved RNA fragments were primed into random hexamers into first strand cDNA using reverse transcriptase and random primers. The RNA template removed and a replacement strand synthesized, incorporating dUTP in place of dTTP to generate ds cDNA. These cDNA fragments then had the addition of a single 'A' base and subsequent ligation of the adapter. The products were then purified and enriched with PCR to create the final cDNA library. Paired-end strand-specific RNA-seq
Experiment attributes:
GEO Accession: GSM3381512
Links:
Runs: 1 run, 33.3M spots, 6.7G bases, 3.3Gb
Run# of Spots# of BasesSizePublished
SRR781372933,274,0846.7G3.3Gb2019-01-28

ID:
6323444

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