Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1081877 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:We optimzed ATAC-seq library preparation for use with Drosophila melanogaster. The protocol addresses factors specific to fruit flies, such as the insect exoskeleton and smaller genome size. The optimized protocol provides guidelines for sample input, nuclei isolation, and enzymatic reaction times. The data included here were generated using our optimized library preparation workflow.
2022-04-20 | GSE197760 | GEO
Project description:We evaluated the effect of the small RNA library preparation method on 5' tRNA-halves and miRNA abundance in libraries prepared from serum RNA using three commercially available small RNA library preparation kits (TruSeq small RNA library preparation kit v2 (Illumina), TailorMix miRNA sample preparation kit v2 (Seqmatic) and the NEBNext Multiplex Small RNA library prep kit (New England Biolabs)). RNA isolated from 100 µl of serum collected from healthy mice was used as input for the preparation of a small RNA library in duplicate and libraries were single end sequenced.
2017-02-22 | GSE88914 | GEO
Project description:We report performance of six different protocols for small RNAseq library preparation and of a method utilizing sequencing of probes targeting microRNAs (HTG EdgeSeq). Recently, small RNA sequencing (small RNA-seq) has been introduced as a method for quantifying circulating microRNAs (miRNAs) and enabling their global profiling without prior knowledge of target sequences. Despite its great promise, small RNA-seq has not delivered the expected outcomes, particularly due to ligation and PCR bias introduced within the workflow. In this study, we assessed the performance of all existing approaches to the small RNA-seq of miRNAs in plasma samples: original two adapter ligation approach; single adapter ligation with subsequent circularization; polyadenylation; use of randomized adapters; and use of unique molecular identifiers (UMI). Using comprehensive set of metrics, we evaluated each protocol in terms of yield, precision, accuracy, sensitivity, and ability to detect isomiRs. Moreover, we assessed performance of targeted RNA-seq method utilizing hybridization probes across relevant metrics and together with RT-qPCR we used it as a reference for accuracy evaluation. The best results were delivered by targeted RNA-seq outperforming other methods in all relevant parameters. The protocols using randomized adapters or UMIs showed consistent good performance across all of the assessed metrics. In contrast, the polyadenylation approach generated a high percentage of discarded reads and impeded the analysis of isomiRs. The single adapter ligation with subsequent circularization failed to prevent ligation bias and the traditional two adapter ligation approach achieved the worse scores in the majority of tested metrics. To sum, we provide a comprehensive comparison that can serve as a guide for new users interested in analysis of circulating miRNAs and as a reference for further comparative studies.
2020-12-31 | GSE149513 | GEO
Project description:We extended the investigation of nuclear transcript changes under drought to an overarching characterization of the post(transcriptome), including alternative splicing of nuclear transcripts, accumulation of organellar (chloroplast and mitochondrion) transcripts, editing and splicing of organellar transcripts, and furthermore, accumulation of transposable elements. To this end, long non-coding (lncRNA-Seq) instead of mRNA-Seq was applied, because the lncRNA-Seq workflow employs library preparation after ribosomal RNA depletion instead of library preparation of enriched mRNAs (mRNA-Seq).
2023-08-09 | GSE202931 | GEO
Project description:Here, we compile valuable insights gathered over years of generating Ribo-seq datasets from different plant species and experimental setups. We tested the effects of variable ribonuclease treatments for the generation of ribosome protected fragments (RPFs). We tested rRNA depletion strategies designed specifically for Arabidopsis and Tobacco. We also compare ligation-free to ligation-based library preparation strategies for generating Ribo-seq libraries.
2024-07-05 | GSE226508 | GEO
Project description:Splinted Ligation Adapter Tagging, a novel library preparation for whole genome bisulphite sequencing
| PRJNA350687 | ENA
Project description:Ribosome profiling has emerged as a powerful tool for genome-wide measurements of translation, but library construction requires multiple ligation steps and remains cumbersome relative to more conventional deep sequencing experiments. We report a new approach to ribosome profiling that does not require ligation. Library construction for ligation-free ribosome profiling can be completed in one day with as little as 1 ng of purified RNA footprints. We used ligation-free ribosome profiling to identify new patterns of cell type-specific translation in the brain and tested its ability to identify translational targets of mTOR signaling in the brain.
2016-06-07 | GSE78163 | GEO
Project description:We present a technique that leverages nucleosome disruption methodologies with the widely adopted 10x Genomics ATAC-seq library preparation workflow to produce scWGS profiles for copy number analysis in high throughput. We further demonstarte the ability to leverage indexed tagmentation using off-the-shelf reagents to perform scWGS multiplexing, or a co-assay that produces scWGS+ATAC data fromt he same cells, also witht he ability to multipelx samples.
2023-09-18 | GSE243430 | GEO