Project description:Next-Generation-Sequencing (NGS) technologies have led to important improvement in the detection of new or unrecognized infective agents, related to infectious diseases. In this context, NGS high-throughput technology can be used to achieve a comprehensive and unbiased sequencing of the nucleic acids present in a clinical sample (i.e. tissues). Metagenomic shotgun sequencing has emerged as powerful high-throughput approaches to analyze and survey microbial composition in the field of infectious diseases. By directly sequencing millions of nucleic acid molecules in a sample and matching the sequences to those available in databases, pathogens of an infectious disease can be inferred. Despite the large amount of metagenomic shotgun data produced, there is a lack of a comprehensive and easy-use pipeline for data analysis that avoid annoying and complicated bioinformatics steps. Here we present HOME-BIO, a modular and exhaustive pipeline for analysis of biological entity estimation, specific designed for shotgun sequenced clinical samples. HOME-BIO analysis provides comprehensive taxonomy classification by querying different source database and carry out main steps in metagenomic investigation. HOME-BIO is a powerful tool in the hand of biologist without computational experience, which are focused on metagenomic analysis. Its easy-to-use intrinsic characteristic allows users to simply import raw sequenced reads file and obtain taxonomy profile of their samples.
Project description:The draft genome of L. sativa (lettuce) cv. Tizian was sequenced in two Illumina sequencing runs, mate pair and shotgun. This entry contains the RAW sequencing data.
Project description:The purpose of this study was to measure DNA methylation and siRNA expression across the maize genome. The experimental data was derived from shotgun bisulfite sequencing, siRNA sequencing, and mRNA sequencing (Illumina, single end for all three)
Project description:Whole-genome sequencing is an important way to understand the genetic information, gene function, biological characteristics, and living mechanisms of organisms. There is no difficulty to have mega-level genomes sequenced at present. However, we encountered a hard-to-sequence genome of Pseudomonas aeruginosa phage PaP1. The shotgun sequencing method failed to dissect this genome. After insisting for 10 years and going over 3 generations of sequencing techniques, we successfully dissected the PaP1 genome with 91,715 bp in length. Single-molecule sequencing revealed that this genome contains lots of modified bases, including 51 N6-methyladenines (m6A) and 152 N4-methylcytosines (m4C). At the same time, further investigations revealed a novel immune mechanism of bacteria, by which the host bacteria can recognize and repel the modified bases containing inserts in large scale, and this led to the failure of the shotgun method in PaP1 genome sequencing. Strategy of resolving this problem is use of non-library dependent sequencing techniques or use of the nfi- mutant of E. coli DH5M-NM-1 as the host bacteria to construct the shotgun library. In conclusion, we unlock the mystery of phage PaP1 genome hard to be sequenced, and discover a new mechanism of bacterial immunity in present study. Methylation profiling of Pseudomonas aeruginosa phage PaP1 using kinetic data generated by single-molecule, real-time (SMRT) sequencing on the PacBio RS.
Project description:This dataset consists of two individual sample-multiplexing (MULTI-seq) single-cell RNA sequencing experiments, MB10x01 and MB10x02. Single-cell RNA sequencing (10X Genomics) analyses were performed on a microfluidic 3D in vitro blood-brain-barrier model (containing primary human brain microvascular endothelial cells, brain vascular pericytes, and astrocytes) perfused with P. falciparum egress product (MB10x01) or P. falciparum-infected red blood cells (RBC) (MB10x02). Dataset MB10x01 included two samples multiplexed by MULTI-seq sample barcoding (TCCTCGAA for control RBC lysate, ATGCGATG for P. falciparum egress product). P. falciparum egress product was obtained by letting tightly synchronized P. falciparum-infected RBC egress in media used for perfusions (5x10^7 infected RBC/ml). 3D blood-brain-barrier models perfused with P. falciparum egress products were incubated for 24 hours and compared to a control perfused with uninfected red blood cell lysate. MULTI-seq barcoding (McGinnis et al.Ê2019) was used for sample-barcoding of these two conditions, and the dataset contains cDNA (transcriptome) and sample barcode read files. Dataset MB10x02 included three samples multiplexed by MULTI-seq sample barcoding (GCTATGCA for control RBC, CGATACTG for Trophozoite stage, TACGCAGT for Schizont stage). 3D blood-brain-barrier models were perfused for 30 minutes with P. falciparum-infected RBC in the Trophozoite stage (26-34 hours post invasion) or Schizont stage (42-48 hours post invasion) (5x10^7 infected RBC/ml). After a 20-minute wash, the 3D blood-brain-barrier models were incubated with the bound P. falciparum-infected RBC for 6 hours and compared to uninfected RBC perfused controls. MULTI-seq barcoding was used for sample-barcoding of the three conditions, and the dataset contains cDNA (transcriptome) and sample barcode read files.
Project description:Part of a set of highly integrated epigenome maps for Arabidopsis thaliana. Keywords: Illumina high-throughput bisulfite sequencing Whole genome shotgun bisulfite sequencing of wildtype Arabidopsis plants (Columbia-0), and met1, drm1 drm2 cmt3, and ros1 dml2 dml3 null mutants using the Illumina Genetic Analyzer.