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    Protocols in Current Issue
    Determination of Mn Concentrations in Synechocystis sp. PCC6803 Using ICP-MS
    Authors:  Fabian Brandenburg, Hanan Schoffman, Nir Keren and Marion Eisenhut, date: 12/05/2017, view: 104, Q&A: 0
    Manganese (Mn) is an essential micronutrient for all photoautotrophic organisms. Two distinct pools of Mn have been identified in the cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis), with 80% of the Mn residing in the ...
    Quantification of Bacterial Polyhydroxybutyrate Content by Flow Cytometry
    Authors:  Antonio Lagares (Jr.) and Claudio Valverde, date: 12/05/2017, view: 102, Q&A: 0
    We describe here a detailed protocol for the quantification of the intracellular content of polyhydroxybutyrate (PHB) in a population of bacterial cells by flow cytometry, which is based on a consensus of several previously reported works.
    Stationary-phase Mutagenesis Soft-agar Overlay Assays in Bacillus subtilis
    Elucidating how a population of non-growing bacteria generates mutations improves our understanding of phenomena like antibiotic resistance, bacterial pathogenesis, genetic diversity and evolution. To evaluate mutations that occur in nutritionally ...
    Determination of Mn Concentrations in Synechocystis sp. PCC6803 Using ICP-MS
    Authors:  Fabian Brandenburg, Hanan Schoffman, Nir Keren and Marion Eisenhut, date: 12/05/2017, view: 104, Q&A: 0
    [Abstract] Manganese (Mn) is an essential micronutrient for all photoautotrophic organisms. Two distinct pools of Mn have been identified in the cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis), with 80% of the Mn residing in the periplasm and 20% in cytoplasm and thylakoid lumen (Keren et al., 2002). In this protocol, we ...
    Quantification of Bacterial Polyhydroxybutyrate Content by Flow Cytometry
    Authors:  Antonio Lagares (Jr.) and Claudio Valverde, date: 12/05/2017, view: 102, Q&A: 0
    [Abstract] We describe here a detailed protocol for the quantification of the intracellular content of polyhydroxybutyrate (PHB) in a population of bacterial cells by flow cytometry, which is based on a consensus of several previously reported works.
    Stationary-phase Mutagenesis Soft-agar Overlay Assays in Bacillus subtilis
    [Abstract] Elucidating how a population of non-growing bacteria generates mutations improves our understanding of phenomena like antibiotic resistance, bacterial pathogenesis, genetic diversity and evolution. To evaluate mutations that occur in nutritionally stressed non-growing bacteria, we have employed the strain B. subtilis YB955, which measures ...
    Design and Direct Assembly of Synthesized Uracil-containing Non-clonal DNA Fragments into Vectors by USERTM Cloning
    [Abstract] This protocol describes how to order and directly assemble uracil-containing non-clonal DNA fragments by uracil excision based cloning (USER cloning). The protocol was generated with the goal of making synthesized non-clonal DNA fragments directly compatible with USERTM cloning. The protocol is highly efficient and would be compatible ...
    [Bio101] A Quick and Easy Method for Making Competent Escherichia coli Cells for Transformation Using Rubidium Chloride
    Authors:  Nidhi Sharma, M. Ximena Anleu Gil and Diego Wengier, date: 11/05/2017, view: 343, Q&A: 0
    [Abstract] This protocol describes a quick and efficient method to make competent E. coli cells for transformation using rubidium chloride. Commercial competent cells are expensive and this protocol provides a cheaper alternative to them.
    Crude Preparation of Lipopolysaccharide from Helicobacter pylori for Silver Staining and Western Blot
    Authors:  Hong Li and Mohammed Benghezal, date: 10/20/2017, view: 523, Q&A: 0
    [Abstract] This protocol provides an easy and rapid method to prepare lipopolysaccharide from the gastric pathogen Helicobacter pylori for visualization on acrylamide gels by silver staining and for detecting the presence of Lewis antigens by Western blot. The silver staining is a four-step procedure, involving a 20 min-oxidation step, a 10 ...
    Capturing Z-stacked Confocal Images of Living Bacteria Entering Hydathode Pores of Cauliflower
    Authors:  Aude Cerutti and Alain Jauneau, date: 10/20/2017, view: 793, Q&A: 0
    [Abstract] The present protocol to visualize living bacteria at the pore level of cauliflower hydathodes is simple and trained users in confocal microscopy can execute it successfully. It can be easily adapted to capture images with other plant-microorganism interactions at the leaf surface and should be useful to obtain important information on pore and ...
    Xanthomonas oryzae pv. oryzae Inoculation and Growth Rate on Rice by Leaf Clipping Method
    Authors:  Yinggen Ke, Shugang Hui and Meng Yuan, date: 10/05/2017, view: 681, Q&A: 0
    [Abstract] Bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the most serious bacterial diseases and a major impediment to the increase of rice yield. Appropriate methods for inoculation of Xoo and disease scoring are necessary to investigate the nature of the disease and the mechanism of plant ...
    Protocol for Construction of a Tunable CRISPR Interference (tCRISPRi) Strain for Escherichia coli
    Authors:  Xin-tian Li, Cindy Sou and Suckjoon Jun, date: 10/05/2017, view: 917, Q&A: 0
    [Abstract] We present a protocol for construction of tunable CRISPR interference (tCRISPRi) strains for Escherichia coli. The tCRISPRi system alleviates most of the known problems of plasmid-based expression methods, and can be immediately used to construct libraries of sgRNAs that can complement the Keio collection by targeting both essential and ...
    Quantification of Densities of Bacterial Endosymbionts of Insects by Real-time PCR
    Author:  Daisuke Kageyama, date: 10/05/2017, view: 552, Q&A: 0
    [Abstract] Increased attention has been paid to the endosymbiotic bacteria of insects. Because most insect endosymbionts are uncultivable, quantitative PCR (qPCR) is a practical and convenient method to quantify endosymbiont titers. Here we report a protocol for real-time qPCR based on SYBR Green I fluorescence as well as some tips to prevent possible ...