Protocols in Current Issue
    Assembly of a Custom-made Device to Study Spreading Patterns of Pseudomonas putida Biofilms
    Authors:  David R. Espeso, Esteban Martínez-García and Víctor de Lorenzo, date: 05/20/2019, view: 1103, Q&A: 0
    [Abstract] Biofilms are bacterial communities in the shape of exopolysaccharide matrix-encased aggregates attached onto interphases able to resist environmental aggressions. The development of bacteria in the shape of biofilms deeply affects the performance of many industrial processes which work with fluidic systems, where bacteria may settle and prosper. ...
    Microfluidics-Based Analysis of Contact-dependent Bacterial Interactions
    Authors:  Robert Cooper, Lev Tsimring and Jeff Hasty, date: 08/20/2018, view: 2286, Q&A: 0
    [Abstract] Bacteria in nature live in complex communities with multiple cell types and spatially-dependent interactions. Studying cells in well-mixed environments such as shaking culture tubes or flasks cannot capture these spatial dynamics, but cells growing in full-fledged biofilms are difficult to observe in real time. We present here a protocol for ...
    Quantification of the Composition Dynamics of a Maize Root-associated Simplified Bacterial Community and Evaluation of Its Biological Control Effect
    Authors:  Ben Niu and Roberto Kolter, date: 06/20/2018, view: 3142, Q&A: 0
    [Abstract] Besides analyzing the composition and dynamics of microbial communities, plant microbiome research aims to understanding the mechanism of plant microbiota assembly and their biological functions. Here, we describe procedures to investigate the role of bacterial interspecies interactions in root microbiome assembly and the beneficial effects of the ...
    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: 5851, 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 ...
    A Modified Low-quantity RNA-Seq Method for Microbial Community and Diversity Analysis Using Small Subunit Ribosomal RNA
    Authors:  Yong-Wei Yan, Ting Zhu, Bin Zou and Zhe-Xue Quan, date: 05/05/2018, view: 2127, Q&A: 0
    [Abstract] We propose a modified RNA-Seq method for small subunit ribosomal RNA (SSU rRNA)-based microbial community analysis that depends on the direct ligation of a 5’ adaptor to RNA before reverse-transcription. The method requires only a low-input quantity of RNA (10-100 ng) and does not require a DNA removal step. Using this method, we could obtain more ...
    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 ...
    Tagged Highly Degenerate Primer (THDP)-PCR for Community Analysis of Methane- and Ammonia-oxidizing Bacteria Based on Copper-containing Membrane-bound Monooxygenases (CuMMO)
    Authors:  Jian-Gong Wang, Fei Xia, Jemaneh Zeleke, Bin Zou and Zhe-Xue Quan, date: 06/20/2017, view: 4027, Q&A: 0
    [Abstract] We describe a two-step PCR strategy using tagged highly degenerate primer (THDP-PCR) targeting copper-containing membrane-bound monooxygenases (CuMMO) genes for community analysis of methane- or ammonia-oxidizing bacteria. This strategy consists of a primary CuMMO gene-specific PCR followed by a secondary PCR with a tag as a single primer. This ...

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