Project description:Purpose: We want to know whether Ino80C contribute to chromatin silencing at both euchromatin and heterochromatin Methods: All yeast cells were collected in exponential phase. Total RNA was extracted with phenol, digested with DNase and further purified by Trizol. Libraries of mRNA were prepared with Illumina TruSeq RNA Sample Prep Kits v2 or stranded RNA Sample Prep Kits. Libraries were sequenced on Illumina HiSeq 2000. ChIP DNA were purified through chromatin immunoprecipitation using an Arp5, H3K79me3 and H3K4me3 antibody. Libraries were prepared with a KAPA LTP kit and sequenced using the Illumina HiSeq 2000 platform. Conclusion: Our ChIP-Seq and mRNA-Seq data show that Ino80C contributes to silent chromatin in both euchromatin an heterochromatin
Project description:Epithelial (CD31-CD45-EpCAM+) and stromal (CD31-CD45-EpCAM-) lung cells were derived by FACS from ShhCre;Ezh2fl/fl and control ShhCre;Ezh2fl/+ mouse embryos at day E16.5 (3 biological replicates per genotype-tissue combination). Total RNA extracted from the samples was subjected to NGS library preparation using TruSeq Stranded Total RNA with Ribo-Zero (Illumina). Completed libraries from different samples were sequenced on HiSeq 2000 TruSeq with SBS Kit v3 - HS reagents (Illumina) as 100 bp single end reads at the Australian Genome Research Facility.
Project description:RNA-sequencing analysis from whole heart ribosome depleted RNA from the 2-week old WT and Lmna-/- mice (N=5) . Strand specific RNA seq libraries where prepared form ribosome-depleted cardiac RNA samples using the Illumina TruSeq stranded total RNA library preparation kit. The samples weresequenced on the Illumina HiSeq 4000 instrument using the paired-end sequencing reagents to generate100 base paired end reads.
Project description:RNA-seq experiment to study the role of Pat1b in HEK293T cells. HEK293T cells were treated twice with siRNA with Lipofectamine 2000 then harvested after 48hours. Total RNA was extracted with TriReagent. Ribo-Zero TruSeq stranded mRNA libraries were prepared for each sample and sequenced on Illumina NextSeq 500 Sequencing System providing around 100 million reads per sample (around 75 bp paired-end reads).
Project description:In total, 70 samples on macroscopically preserved and lesioned OA cartilage from the same patient was taken for RNA-seq. Subsequently, paired-end 2×100 bp RNA library sequencing (Illumina TruSeq RNA Library Prep Kit, Illumina HiSeq 2000 and TruSeq Stranded Total RNA LT Sample Prep Kit, Illumina HiSeq 4000) was performed.
Project description:In the present study, RNA sequencing of 6 human keratinocytes cell samples was performed. They are 3 samples of Poly (I:C)-treated cells, and 3 samples from un-treated control cells, from 3 different human donors. Total RNA was isolated from the cell samples and one RNA sequencing library was prepared from the 6 RNA samples with the Illumina TruSeq Stranded mRNA HT technolog. This approach uses fragmentation, a poly-T oligo pulldown and sequencing adaptor ligation. RNA sequencing was performed on the Illumina NextSeq500 next generation sequencing system and its high output mode with 1x75 bp single-end red chemistry.
Project description:IPF (n=20) and control (n=19) samples were obtained through the LTRC and were sequenced on an Illumina HiSeq 2000 following TruSeq RNA Sample Prep Kit v2 library preparation.
Project description:Library preparation is a key step in gene expression quantification. There are considerable advantages to both strand specific sequencing and the ability to sequence samples with very small amounts of starting material. Until recently there was no kit available that allowed both simultaneously. The standard Illumina-TruSeq stranded mRNA Sample Preparation kit requires abundant starting quantity while the Takara Bio-SMART-Seq® v4 Ultra® Low Input RNA kit allows for ultra low starting quantities but sacrifices strand specificity. Recently a kit that can do both, SMARTer® Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian by Takara Bio, has become available. Evaluating the performance and effects of these sample preparation kits is a critical determinant for selecting the appropriate sequencing protocol, but a comprehensive comparison is currently missing. To address this we performed a detailed comparative analysis of sequencing libraries prepared with the three kits. We prepared a set of samples representing two experimental conditions with each kit, allowing for comparison of the kits in a standard realistic differential expression analysis. We find substantial differences in the levels of alignment and differential gene expression. Using differential expression analysis we show that using Pico results in identifying 55% less differentially expressed genes than TruSeq. Nevertheless, using gene pathway enrichment analysis we find similar results for all three kits, indicating that ultimately comparable functional results can be reached.
Project description:Epithelial (CD31-CD45-EpCAM+) and stromal (CD31-CD45-EpCAM-) lung cells were derived by FACS from ShhCre;Ezh2fl/fl and control ShhCre;Ezh2fl/+ mouse embryos at day E16.5 (3 biological replicates per genotype-tissue combination). Total RNA extracted from the samples was subjected to NGS library preparation using TruSeq Stranded Total RNA with Ribo-Zero (Illumina). Completed libraries from different samples were sequenced on HiSeq 2000 TruSeq with SBS Kit v3 - HS reagents (Illumina) as 100 bp single end reads at the Australian Genome Research Facility. RNA-seq gene expression profiles from Ezh2 deficient and control lung epithelial and stromal cell populations at day E16.5 (3 replicates per genotype-tissue combination).