Systems Biology

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    Protocols in Current Issue
    Extraction and 16S rRNA Sequence Analysis of Microbiomes Associated with Rice Roots
    Authors:  Joseph Edwards, Christian Santos-Medellín and Venkatesan Sundaresan, date: 06/20/2018, view: 6137, Q&A: 0
    [Abstract] Plant roots associate with a wide diversity of bacteria and archaea across the root-soil spectrum. The rhizosphere microbiota, the communities of microbes in the soil adjacent to the root, can contain up to 10 billion bacterial cells per gram of soil (Raynaud and Nunan, 2014) and can play important roles for the fitness of the host plant. Subsets ...
    Brief Protocol for EDGE Bioinformatics: Analyzing Microbial and Metagenomic NGS Data
    [Abstract] Next-generation sequencing (NGS) offers unparalleled resolution for untargeted organism detection and characterization. However, the majority of NGS analysis programs require users to be proficient in programming and command-line interfaces. EDGE bioinformatics was developed to offer scientists with little to no bioinformatics expertise a ...
    A Method to Convert mRNA into a Guide RNA (gRNA) Library without Requiring Previous Bioinformatics Knowledge of the Organism
    Author:  Hiroshi Arakawa, date: 05/20/2017, view: 5385, Q&A: 0
    [Abstract] While the diversity of species represents a diversity of special biological abilities, many of the genes that encode those special abilities in a variety of species are untouched, leaving an untapped gold mine of genetic information; however, despite current advances in genome bioinformatics, annotation of that genetic information is incomplete in ...
    Next-generation Sequencing of the DNA Virome from Fecal Samples
    Authors:  Cynthia L. Monaco and Douglas S. Kwon, date: 03/05/2017, view: 5687, Q&A: 0
    [Abstract] Herein we describe a detailed protocol for DNA virome analysis of low input human stool samples (Monaco et al., 2016). This protocol is divided into four main steps: 1) stool samples are pulverized to evenly distribute microbial matter; 2) stool is enriched for virus-like particles and DNA is extracted by phenol-chloroform; 3) purified ...
    Sequencing of Ebola Virus Genomes Using Nanopore Technology
    Author:  Thomas Hoenen, date: 11/05/2016, view: 6647, Q&A: 0
    [Abstract] Sequencing of virus genomes during disease outbreaks can provide valuable information for diagnostics, epidemiology, and evaluation of potential countermeasures. However, particularly in remote areas logistical and technical challenges can be significant. Nanopore sequencing provides an alternative to classical Sanger and next-generation ...
    Chromosome Dosage Analysis in Plants Using Whole Genome Sequencing
    Authors:  Ek Han Tan, Luca Comai and Isabelle M. Henry, date: 07/05/2016, view: 6823, Q&A: 0
    [Abstract] Relative chromosome dosage, i.e., increases or decreases in the number of copies of specific chromosome regions in one sample versus another, can be determined using aligned read-counts from Illumina sequencing (Henry et al., 2010). The following protocol was used to identify the different classes of aneuploids that ...
    Protocol for the Generation of a Transcription Factor Open Reading Frame Collection (TFome)
    [Abstract] The construction of a physical collection of open reading frames (ORFeomes) for genes of any model organism is a useful tool for the exploration of gene function, gene regulation, and protein-protein interaction. Here we describe in detail a protocol that has been used to develop the first collection of transcription factor (TF) and co-regulator ...
    Human, Bacterial and Fungal Amplicon Collection and Processing for Sequencing
    Author:  Julia Oh, date: 05/20/2015, view: 8132, Q&A: 3
    [Abstract] Sequencing taxonomic marker genes is a powerful tool to interrogate the composition of microbial communities. For example, bacterial and fungal community composition can be evaluated in parallel using the 16S ribosomal RNA gene for bacteria or the internal transcribed spacer region in fungi. These are conserved regions that are universal to a ...
    Determining the Relative Fitness Score of Mutant Viruses in a Population Using Illumina Paired-end Sequencing and Regression Analysis
    Authors:  Hangfei Qi, C. Anders Olson, Nicholas C. Wu, Yushen Du and Ren Sun, date: 05/20/2015, view: 5929, Q&A: 0
    [Abstract] Recent advances in DNA sequencing capacity to accurately quantify the copy number of individual variants in a large and diverse population allows in parallel determination of the phenotypic effects caused by each genetic modification. This systematic profiling approach is a combination of forward and reverse genetics, which we refer to as ...
    siRNA Screening for Genes Involved in HSV-1 Replication
    Authors:  Samantha J. Griffiths and Jürgen Haas, date: 08/20/2014, view: 7240, Q&A: 0
    [Abstract] Small interfering RNAs (siRNAs) are small (typically 18-24 nucleotides) RNA molecules capable of silencing gene expression post-transcriptionally and as such, they provide a simple method by which the role of individual genes in complex cellular systems can be easily assessed. As siRNAs are easy to use experimentally, and can be designed to target ...



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