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Isolation and Culture of Single Myofiber and Immunostaining of Satellite Cells from Adult C57BL/6J Mice

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Quantitative Transformation Efficiency Assay for Bacillus subtilis

Authors: Christian L. Loyo and Briana M. Burton, date: 12/05/2018, view: 1420, Q&A: 0

[Abstract] Bacillus subtilis (B. subtilis) is a model Gram-positive organism used to study a variety of physiological states and stress responses, one of which is the development of competence. Competence refers to the physiological state of a cell which allows it to be transformed naturally. Through induction of competence, the efficiency ...

Extraction and Analysis of Bacterial Teichoic Acids

Authors: Kelvin Kho and Timothy C. Meredith, date: 11/05/2018, view: 1509, Q&A: 0

[Abstract] Teichoic acids (TA) are anionic polymers comprised of polyol phosphate repeat units that are abundant in the cell wall of Gram-positive bacteria. Both wall teichoic acid (WTA) and lipoteichoic acid (LTA) play important roles in regulating cell wall remodeling as well as conferring antibiotic resistance. To analyze TA, we describe a polyacrylamide ...

Extraction and Quantification of Polyphosphate (polyP) from Gram-negative Bacteria

Authors: Jan-Ulrik Dahl, Lihan Xie and Ursula Jakob, date: 09/20/2018, view: 1426, Q&A: 0

[Abstract] Polyphosphate (polyP), a universally conserved biomolecule, is composed of up to 1,000 phosphate monomers linked via phosphoanhydride bonds. Reaching levels in bacteria that are in the high nmoles per mg protein range, polyP plays important roles in biofilm formation and colonization, general stress protection and virulence. Various protocols for ...

Selective Isolation of Retroviruses from Extracellular Vesicles by Intact Virion Immunoprecipitation

Authors: Tyler Milston Renner, Kasandra Bélanger and Marc-André Langlois, date: 09/05/2018, view: 1433, Q&A: 0

[Abstract] There exists a wide variety of techniques to isolate and purify viral particles from cell culture supernatants. However, these techniques vary greatly in ease of use, purity, yield and impact on viral structural integrity. Most importantly, it is becoming evident that secreted extracellular vesicles (EVs) co-purify with retroviruses using nearly ...

Quantification of Hydrogen Sulfide and Cysteine Excreted by Bacterial Cells

Authors: Sergey Korshunov and James A. Imlay, date: 05/20/2018, view: 2032, Q&A: 0

[Abstract] Bacteria release cysteine to moderate the size of their intracellular pools. They can also evolve hydrogen sulfide, either through dissimilatory reduction of oxidized forms of sulfur or through the deliberate or inadvertent degradation of intracellular cysteine. These processes can have important consequences upon microbial communities, because ...

Evaluation of the Condition of Respiration and Photosynthesis by Measuring Chlorophyll Fluorescence in Cyanobacteria

Authors: Takako Ogawa and Kintake Sonoike , date: 05/05/2018, view: 2363, Q&A: 0

[Abstract] Chlorophyll fluorescence measurements have been widely used to monitor the condition of photosynthesis. Furthermore, chlorophyll fluorescence from cyanobacteria reflects the condition of respiration, since cyanobacterial photosynthesis shares several components of electron transport chain with respiration. This protocol presents the method to ...

Determination of Intracellular Osmolytes in Cyanobacterial Cells

Authors: Xiaoming Tan, Kuo Song, Cuncun Qiao and Xuefeng Lu, date: 04/20/2018, view: 1838, Q&A: 0

[Abstract] Most of the cyanobacteria accumulate osmolytes including sucrose, glucosylglycerol, in their cells in response to salt stress. Here we describe a protocol of our laboratory for extraction and quantification of cyanobacterial intracellular sucrose and glucosylglycerol. We have confirmed this protocol was applicable to at least four kinds of ...

Bacterial Competition Assay Based on Extracellular D-amino Acid Production

Authors: Laura Alvarez and Felipe Cava, date: 04/05/2018, view: 2987, Q&A: 0

[Abstract] Bacteria live in polymicrobial communities under tough competition. To persist in a specific niche many species produce toxic extracellular effectors as a strategy to interfere with the growth of nearby microbes. One of such effectors are the non-canonical D-amino acids. Here we describe a method to test the effect of D-amino acid production in ...

ROS Detection in Botryococcus braunii Colonies with CellROX Green Reagent

Authors: Edmundo Lozoya-Gloria, Ivette Cornejo-Corona, Hem R. Thapa, Daniel R. Browne and Timothy P. Devarenne, date: 08/20/2017, view: 3655, Q&A: 0

[Abstract] We analyzed the reactive oxygen species (ROS) accumulation in the colony-forming green microalga Botryococcus braunii in response to several stress inducers such as NaCl, NaHCO₃, salicylic acid (SA), methyl jasmonate, and acetic acid. A staining assay using the fluorescent dye CellROX Green was used. CellROX Green is a ...

Analysis of N-acetylmuramic acid-6-phosphate (MurNAc-6P) Accumulation by HPLC-MS

Authors: Marina Borisova and Christoph Mayer, date: 08/05/2017, view: 3943, Q&A: 0

[Abstract] We describe here in detail a high-performance liquid chromatography-mass spectrometry (HPLC-MS)-based method to determine N-acetylmuramic acid-6-phosphate (MurNAc-6P) in bacterial cell extracts. The method can be applied to both Gram-negative and Gram-positive bacteria, and as an example we use Escherichia coli cells in this ...

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