34e0c98e-d7b0-4641-b82f-b8f60bcc1b1c - samples
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ABSTRACT:
PROVIDER: EGAD00001010309 | EGA |
REPOSITORIES: EGA
Similar Datasets
Project description:To gain insights how loss of PIK3R1 confers resistance to pharmacological Notch inhibition in T-ALL cells, we performed gene expression analysis. Libraries for mRNA-seq were prepared with the Stranded mRNA Ligation method (Illumina) starting from 1µg RNA, according to manufacturer’s instructions. Libraries, all bearing unique dual indices, were subsequently loaded at 250pM in a HiSeq 4000 instrument (Illumina) and sequenced according to manufacturer’s instructions, yielding paired-end reads of 75 nucleotides. Reads were trimmed of their adapters with bcl2fastq v2.20 (Illumina) and quality-controlled with FastQC v0.11.9. PIK3R1 deficiency leads to up-regulation of cell cycle and pro-survival genes in response to Notch inhibition.
2023-06-26 | GSE221577 | GEO
Project description:Purpose: The aim of this study is (1) to identify the chromatin occupancy of the epigenetic regulator Smchd1 in neural stem cells (NSCs) derived from E14.5 mouse brain; (2) to profile key epigenetic marks H3K4me3, H3K27me3 and DNA methylation in wild type and Smchd1 null NSCs; (3) to identify the chromatin occupancy of Ctcf in wild type and Smchd1 null NSCs. Methods: Chromatin immunoprecipitation for Smchd1, H3K4me3, H3K27me3 and Ctcf was performed essentially as in (Nelson et al. 2006). Briefly, nuclei were isolated from formaldehyde crosslinked NSCs and chromatin was fragmented by sonication. Chromatin immunoprecipitation was performed with corresponding antibodies for Smchd1, H3K4me3 and H3K27me3. DNA was extracted from the immunoprecipitated fraction following reverse-crosslinking. Isolated DNA was used to generate sequencing libraries with Illumina's TruSeq DNA Sample Preparation Kit or Ovation Ultralow system (NuGen) according to manufacturer's instruction. Libraries were pooled and sequenced on the Illumina HiSeq 2000 platform for 100 bp single-end reads. Image analysis was performed in real time by the HiSeq Control Software (HCS) v1.4.8 and Real Time Analysis (RTA) v1.12.4.2, running on the instrument computer. Real-time base calling on the HiSeq instrument computer was performed with the RTA software. Illumina CASAVA1.8 pipeline was used to generate the sequence data. To examine the level of DNA methylation, genomic DNA was extracted using an AllPrep DNA/RNA Mini Kit (Qiagen) and methylated DNA was isolated via binding to the methyl-CpG binding domain of human MBD2 protein coupled beads using the MethylMiner methylated DNA enrichment kit (Life Technologies) according to the manufacturer’s instructions. Isolated DNA was used to generate sequencing libraries as for the ChIP-seq experiment with Illumina’s TruSeq DNA Sample Preparation Kit according to manufacturer's instruction and sequenced on the Illumina HiSeq 2000 platform for 49 bp single-end reads. Sequencing analysis was performed as described for the ChIP-seq experiments.
2015-06-11 | GSE65748 | GEO
Project description:To better understand the OLIG2 binding site in the whole-genome of mouse neural stem cells, ChIP was performed using 107 mouse neural stem cells. The ChIP-Seq library was constructed by using DNA SMART ChIP-Seq Kit according to the manufacturer's instructions (Clontech) and was sequenced on the Illumina HiSeq 2000 Sequencer.
2015-11-04 | GSE74646 | GEO
Project description:Abstract: Wallerian degeneration (WD) is a process of autonomous distal degeneration of axons upon injury. Macrophages (MP) of the peripheral nervous system (PNS) are main cellular agent controlling this process. Some evidences suggest that resident PNS-MP along with MP of hematogenous origin may be involved but whether these two subsets exert distinct functions is unknown. Combining MP-designed fluorescent reporters mice, and coherent anti-stoke raman scattering (CARS) imaging of the sciatic nerve, we deciphered the spatio-temporal choreography of resident and recently recruited MP after injury and unveiled distinct functions of these subsets with recruited MP responsible of efficient myelin stripping and clearance while resident MP were involved in axonal regrowth. This work provides clues to tackle selectively cellular processes involved in neurodegenerative diseases. Methods: (relevant for this GEO dataset): RNAseq of sciatic nerve macrophages from mice after CCI: Sorted cells were lysed in 100µl of RA1/TCEP buffer (NucleoSpin RNA XS, Macherey-Nagel), snap frozen in liquid nitrogen and stored at -80C until RNA extraction. All samples were processed in parallel and RNA extraction (without Carrier RNA but with on-column DNase treatment) was performed according to manufacturer's instructions (NucleoSpin RNA XS, Macherey-Nagel). RNA was eluted in RNAse-free water. Preparation of cDNA libraries for RNAseq was done using the SmartSeq method according to manufacturer's instructions (SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing, Clontech/TaKaRa). Due to the low number of cells, the total amount of eluted RNA was used as starting material for reverse transcription, followed by 18 cycles of pre-amplification. 1ng of cDNA were used for RNAseq sequencing library preparation, according to manufacturer's instructions (Nextera XT DNA Library Preparation, Illumina). Final samples pooled library prep were sequenced on a Nextseq 500 ILLUMINA with MidOutPut cartridge (2x130Millions of 75 bases reads) with 2 runs (4plex and 5plex), corresponding to 2x30Millions of (paired-end) reads per sample after demultiplexing.
2020-06-30 | GSE152206 | GEO
Project description:To develop gene-miRNA and pathway-miRNA networks in trastuzumab treatment, we performed a recent miRNA microarray profiling in trastuzumab treated/untreated SKBR3 and BT474 cell lines. The cells were plated at a starting density of 2 millions in 100 mm cell culture dishes and then, they were treated with 6 ug/mL trastuzumab and PBS in two replicates. Total RNA was isolated with the TRIzol reagent (Invitrogen) according to the manufacturer's instructions.
2017-09-21 | GSE104076 | GEO
Project description:Purpose:To understand the change in transcriptome for the overxepression of ΔCSP in HepG2 cells. Methods:Total RNAs of ΔCSP stable expressing or control HepG2 cells were extracted using TRIzol (Invitrogen), following the manufacturer's instructions. RNA-seq and bioinformatic data analysis were performed by Shanghai Sangon Bio Ltd. Briefly, sequencing libraries were generated using VAHTSTM mRNA-seq V2 Library Prep Kit for Illumina® (Vazyme Biotech, Nanjing, China) following manufacturer's recommendations and were sequenced on HiSeq XTen sequencers (Illumina, San Diego, CA). Raw reads in FASTQ format were subjected to quality control using FastQC. RNA-seq reads were aligned to the reference genome using Bowtie. Uniquely mapped reads were used for further analysis. Gene expression levels are expressed as TPM (Transcripts per million reads) and differences in gene expression were calculated with rSeq. Results:There were 492 genes differentially expressed in ΔCSP-overexpressing HepG2 cells compare to the control group respectively (fold change >2 or < 0.5).
2020-12-31 | GSE129323 | GEO
Project description:Purpose: Identification of glucocorticoid recepor binding sites on human kidney proximal tubular cells to better understand epigenetic alterations in diabetic kidney diseases Methods: CUT&RUN assay libraries for cultured cells or human kidneys were generated with the CUTANA kit (EpiCypher, 14-1048). 500,000 cells were mixed and incubated with Concanavalin A (ConA) conjugated paramagnetic beads. Antibodies to the glucocorticoid receptor were added to target samples and IgG was added to negative controls. The remaining steps were performed according to manufacturer’s instructions. Library preparation was performed using the NEBNext Ultra II DNA Library Prep Kit for Illumina (New England BioLabs, E7645S) with manufacturer’s instructions, including minor modifications indicated by CUTANA described above. CUT&RUN libraries were sequenced on a NovaSeq instrument (Illumina, 150 bp paired-end reads). For bulk ATAC-seq, 50,000 nuclei were transposed in 25 μL of ATAC-seq transposition mix [12.5 μL 2× Illumina Tagment DNA (TD) buffer; 10.5 μL nuclease-free water; 2.0 μL Tn5 transposase (Illumina/Nextera, FC-121-1030)] and incubated at 37°C for 1 h on a thermomixer. The transposed DNA was purified with QIAGEN MinElute kit (28004) and amplified with dual indexes.
2022-07-28 | GSE195443 | GEO
Project description:Purpose: To compare the coding and non-coding transcriptomic changes in human gliomas compared with normal brain tissues. Methods: Biopsies were collected from 85 adult glioblastomas, 18 lower grade gliomas, and 15 normal brain tissues. Total RNA was isolated using the Qiagen RNeasy Mini kit or Trizol reagent according to the manufacturer's instruction. Stranded, rRNA-deleted libraries were prepared using the Illumina stranded Total RNA TruSEQ kit according to the manufacturer's instruction. Sequencing was performed on Illumina HiSeq 4000 instrument according to the manufacturer instructions. Reads were aligned to the human genome GRCh38 using HISAT2 (Version 2.1.0). Read counts for each gene were quantified using HTSeq-count (Version 0.10.0) and then normalized by DESeq2 (r package, version 1.20.0). Results: Approximately 20 million paired-end reads at the length of 50 bp were obtained from each sample. The genome mapping efficiency was 85% on average for all samples. Altered expression of 6,132 protein-coding genes and 1,270 lncRNAs were identified. Conclusions:We report the coding and non-coding transcriptomic changes in human gliomas compared with normal brain tissues.
2021-01-26 | GSE147352 | GEO
Project description:The larvae were treated with 200 μM neomycin to deplete the hair cells in neuromast and ampullary organ. Two kinds of sensory epithelia at the ventral side of head were separated at 12 hour-post treatment (hpt) and 24 hpt, respectively. Mechano- and electro- sensory epithelia before neomycin treating were collected respectively as control (untreated).The epithelia near but without two kinds of sensors (EPs) were also collected. Total RNAs were extracted by TRIzol (Invitrogen) using standard protocols. RNA libraries were prepared by TruSeq RNA Sample Prep Kit (Illumina), following the manufacturer's instructions.
2020-12-15 | GSE151096 | GEO
Project description:We report the result of lncRNAs binding to Hes1 in colon cancer SW480 cells using Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Millipore, MD, USA) following the manufacturer's instructions, the RNAs were analyzed by Illumina HiSeq 2500 Sequencing.
2019-07-20 | GSE134551 | GEO