Computational Analysis and Phylogenetic Clustering of SARS-CoV-2 Genomes
作者:Bani Jolly and Vinod Scaria日期:04/20/2021,浏览量:1353,Q&A: 0

COVID-19, the disease caused by the novel SARS-CoV-2 coronavirus, originated as an isolated outbreak in the Hubei province of China but soon created a global pandemic and is now a major threat to

Quantitative Characterization of the Amount and Length of (1,3)-β-D-glucan for Functional and Mechanistic Analysis of Fungal (1,3)-β-D-glucan Synthase
作者:Abhishek Chhetri, Anna Loksztejn and Kenichi Yokoyama日期:04/20/2021,浏览量:993,Q&A: 0

(1,3)-β-d-Glucan synthase (GS) is an essential enzyme for fungal cell wall biosynthesis that catalyzes the synthesis of (1,3)-β-d-glucan, a major and vital component of the cell wall. GS is

Screening for Lysogen Activity in Therapeutically Relevant Bacteriophages
作者:Fernando L. Gordillo Altamirano and Jeremy J. Barr日期:04/20/2021,浏览量:1381,Q&A: 1

Lysogenic phages can integrate into their bacterial host’s genome, potentially transferring any genetic information they possess including virulence or resistance genes, and are therefore

A Sensitive and Specific PCR-based Assay to Quantify Hepatitis B Virus Covalently Closed Circular (ccc) DNA while Preserving Cellular DNA
作者:Benno Zehnder, Stephan Urban and Thomas Tu日期:04/20/2021,浏览量:867,Q&A: 0

Hepatitis B virus (HBV) is the major cause of liver diseases and liver cancer worldwide. After infecting hepatocytes, the virus establishes a stable episome (covalently closed circular DNA, or

Optimized Recombinant Production of Secreted Proteins Using Human Embryonic Kidney (HEK293) Cells Grown in Suspension

Recombinant proteins are an essential milestone for a plethora of different applications ranging from pharmaceutical to clinical, and mammalian cell lines are among the currently preferred systems to

Liposomal Clodronate-mediated Macrophage Depletion in the Zebrafish Model
作者:Linlin Yang, Alison M. Rojas and Celia E. Shiau日期:03/20/2021,浏览量:1143,Q&A: 0

The ability to conduct in vivo macrophage-specific depletion remains an effective means to uncover functions of macrophages in a wide range of physiological contexts. Compared to the murine model,

Multilayered Fabrication Assembly Technique to Engineer a Corneal Stromal Equivalent
作者:Julia Fernández-Pérez, Peter W. Madden and Mark Ahearne日期:03/20/2021,浏览量:1028,Q&A: 0

Tissue engineering has emerged as a strategy to combat the donor shortage of human corneas for transplantation. Synthetic corneal substitutes are currently unable to support the normal phenotype of

Intestinal Co-culture System to Study TGR5 Agonism and Gut Restriction
作者:Snehal N. Chaudhari and A. Sloan Devlin日期:03/20/2021,浏览量:1157,Q&A: 0

The activation of the Takeda G-protein receptor 5 (TGR5, also known as the G protein-coupled bile acid receptor 1, GPBAR1) in enteroendocrine L-cells results in secretion of the anti-diabetic hormone

Retention Using Selective Hooks (RUSH) Cargo Sorting Assay for Live-cell Vesicle Tracking in the Secretory Pathway Using HeLa Cells
作者:Mehrshad Pakdel, Natalia Pacheco-Fernandez and Julia von Blume日期:03/20/2021,浏览量:1075,Q&A: 0

More than 30% of the total amount of proteins synthesized in mammalian cells follow the secretory pathway in order to mature and be properly sorted to their final destinations. Among several

Colorimetric RT-LAMP and LAMP-sequencing for Detecting SARS-CoV-2 RNA in Clinical Samples
作者:Konrad Herbst, Matthias Meurer, Daniel Kirrmaier, Simon Anders, Michael Knop and Viet Loan Dao Thi日期:03/20/2021,浏览量:1204,Q&A: 0

During pandemics, such as the one caused by SARS-CoV-2 coronavirus, simple methods to rapidly test large numbers of people are needed. As a faster and less resource-demanding alternative to detect

Murine Monocyte and Macrophage Culture
作者:Simone M. Haag and Aditya Murthy日期:03/20/2021,浏览量:2273,Q&A: 0

Myeloid progenitors in the bone marrow generate monocytes, macrophages, granulocytes and most dendritic cells. Even though these innate immune cells are part of the same lineage, each cell type plays

Rapid Genome Engineering of Pseudomonas Assisted by Fluorescent Markers and Tractable Curing of Plasmids
作者:Daniel C. Volke, Nicolas T. Wirth and Pablo I. Nikel日期:02/20/2021,浏览量:2213,Q&A: 0

Precise genome engineering has become a commonplace technique for metabolic engineering. Also, insertion, deletion and alteration of genes and other functional DNA sequences are essential for

Expression and Purification of Yeast-derived GPCR, Gα and Gβγ Subunits for Structural and Dynamic Studies
作者:Wenjie Zhao, Xudong Wang and Libin Ye日期:02/20/2021,浏览量:1828,Q&A: 0

In the last several years, as evidence of a surged number of GPCR-G complex structures, the expressions of GPCRs and G proteins for structural biology have achieved tremendous successes, mostly in

Evaluation of the Sequence Variability within the PCR Primer/Probe Target Regions of the SARS-CoV-2 Genome
作者:Kashif Aziz Khan and Peter Cheung日期:12/20/2020,浏览量:1612,Q&A: 0
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; initially named 2019-nCoV) is responsible for the recent coronavirus disease (COVID-19) pandemic, and polymerase chain reaction (PCR) is ...
Native Co-immunoprecipitation Assay to Identify Interacting Partners of Chromatin-associated Proteins in Mammalian Cells
作者:Afzal Husain, Nasim A. Begum, Maki Kobayashi and Tasuku Honjo日期:12/05/2020,浏览量:1949,Q&A: 0

Protein-protein interactions play key roles in nuclear processes including transcription, replication, DNA damage repair, and recombination. Co-immunoprecipitation (Co-IP) followed by western blot or

Agrobacterium-mediated Transformation of Sweet Basil (Ocimum basilicum)
农杆菌介导的罗勒(Ocimum basilicum)转化
作者:Natasha Navet and Miaoying Tian日期:11/20/2020,浏览量:1886,Q&A: 1

Sweet basil (Ocimum basilicum) is a popular herb with high economic value and is currently threatened by a severe oomycete disease. An efficient transformation method is a prerequisite for gene

Colorimetric RT-LAMP Methods to Detect Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)
作者:Gun-Soo Park, Seung-Hwa Baek, Keunbon Ku, Seong Jun Kim, Seung Il Kim, Bum-Tae Kim and Jin-Soo Maeng日期:11/05/2020,浏览量:1947,Q&A: 0
Standard diagnostic methods of Coronavirus Disease 2019 (COVID-19) rely on RT-qPCR technique which have limited point-of-care test (POCT) potential due to necessity of dedicated equipment and ...
Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of Heritable Cas9-free Mutants in Arabidopsis
作者:Yu Jin and Sebastian Marquardt日期:10/20/2020,浏览量:2225,Q&A: 0
CRISPR/Cas9 system directed by a gene-specific single guide RNA (sgRNA) is an effective tool for genome editing such as deletions of few bases in coding genes. However, targeted deletion of ...
Multitarget Immunohistochemistry for Confocal and Super-resolution Imaging of Plant Cell Wall Polysaccharides
作者:Kalina T. Haas, Methieu Rivière, Raymond Wightman and Alexis Peaucelle日期:10/05/2020,浏览量:2285,Q&A: 0
The plant cell wall (PCW) is a pecto-cellulosic extracellular matrix that envelopes the plant cell. By integrating extra-and intra-cellular cues, PCW mediates a plethora of essential physiological ...
A Workflow for Ultra-rapid Analysis of Histone Post-translational Modifications with Direct-injection Mass Spectrometry
作者:Natarajan V. Bhanu, Simone Sidoli and Benjamin A Garcia日期:09/20/2020,浏览量:2357,Q&A: 0
Chromatin modifications, like histone post translational modifications (PTMs), are critical for tuning gene expression and many other aspects of cell phenotype. Liquid chromatography coupled to mass ...
Flow-cytometric Detection of Low-level Reactive Oxygen Species in Cell Lines and Primary Immune Cells
作者:Kevin Bode, Corinna Link, Peter H. Krammer and Heiko Weyd日期:09/05/2020,浏览量:2720,Q&A: 0
Depending on its concentration and cellular origin the production of reactive oxygen species (ROS) in the organism serves a variety of functions. While high concentrations during an oxidative burst ...
In vivo Blood-brain Barrier Permeability Assays Using Clostridium perfringens Epsilon Toxin
作者:Michael R. Mazzucco, Timothy Vartanian and Jennifer R. Linden日期:08/05/2020,浏览量:2316,Q&A: 0
In order for the brain to function properly, a carefully orchestrated homeostasis must be maintained. To help regulate this delicate balance, the brain has developed a highly selective blood-brain ...
Measuring Intracellular Vesicle Density and Dispersion Using Fluorescence Microscopy and ImageJ/FIJI
使用荧光显微镜和ImageJ / FIJI测量细胞内囊泡的密度和分散度
Cell signalling, cell secretion, and plasma membrane repair are processes that critically rely on intracellular vesicles, important components of the endocytic and secretory pathways. More ...
Preparation of Drosophila Polytene Chromosomes, Followed by Immunofluorescence Analysis of Chromatin Structure by Multi-fluorescence Correlations
作者:Terra M. Kuhn, Shawn C. Little and Maya Capelson日期:07/05/2020,浏览量:4337,Q&A: 0
Drosophila larval salivary gland polytene chromosome squashes have been used for decades to analyze genome-wide protein-binding patterns, transcriptional activation processes, and changes in ...
Mouse Adipose Tissue Protein Extraction
作者:Yu A. An and Philipp E. Scherer日期:06/05/2020,浏览量:4070,Q&A: 0
As obesity becomes a global epidemic, the metabolism research field is increasingly focusing on studying the physiological and pathological roles of adipose tissues (AT). However, extracting proteins ...
Confocal and Super-resolution Imaging of RNA in Live Bacteria Using a Fluorogenic Silicon Rhodamine-binding Aptamer
作者:Regina Wirth, Peng Gao, G. Ulrich Nienhaus, Murat Sunbul and Andres Jäschke日期:05/05/2020,浏览量:4087,Q&A: 0
Genetically encoded light-up RNA aptamers have been shown to be promising tools for the visualization of RNAs in living cells, helping us to advance our understanding of the broad and complex life of ...
Unbiased and Tailored CRISPR/Cas gRNA Libraries by Synthesizing Covalently-closed-circular (3Cs) DNA
通过合成共价闭合环状(3Cs)DNA构建无偏和专用CRISPR / Cas gRNA文库
作者:Martin Wegner, Koraljka Husnjak and Manuel Kaulich日期:01/05/2020,浏览量:4445,Q&A: 0
Simplicity, efficiency and versatility of the CRISPR/Cas system greatly contributed to its rapid use in a broad range of fields. Applications of unbiased CRISPR/Cas screenings are increasing and thus ...
Tryptophan Fluorescence Quenching Assays for Measuring Protein-ligand Binding Affinities: Principles and a Practical Guide
作者:Anthony Yammine, Jinlong Gao and Ann H. Kwan日期:06/05/2019,浏览量:12391,Q&A: 0
Tryptophan fluorescence quenching is a type of fluorescence spectroscopy used for binding assays. The assay relies on the ability to quench the intrinsic fluorescence of tryptophan residues within a ...
Analysis of Indole-3-acetic Acid (IAA) Production in Klebsiella by LC-MS/MS and the Salkowski Method
作者:Shraddha Gang, Sheetal Sharma, Meenu Saraf, Martin Buck and Jörg Schumacher日期:05/05/2019,浏览量:15094,Q&A: 1
Many rhizobacteria isolated from plant rhizosphere produce various phytohormones in the form of secondary metabolites, the most common of which is Indole-3-acetic acid (IAA). Here, we detail ...
Single-molecule Fluorescence in situ Hybridization (smFISH) for RNA Detection in Adherent Animal Cells
作者:Gal Haimovich and Jeffrey E. Gerst日期:11/05/2018,浏览量:23242,Q&A: 1
Transcription and RNA decay play critical roles in the process of gene expression and the ability to accurately measure cellular mRNA levels is essential for understanding this regulation. Here, we ...
mRNA Stability Assay Using Transcription Inhibition by Actinomycin D in Mouse Pluripotent Stem Cells
作者:Madara Ratnadiwakara and Minna-Liisa Änkö日期:11/05/2018,浏览量:20659,Q&A: 2
Gene expression is regulated through multiple steps at both transcriptional and post-transcriptional levels. The net abundance of mature mRNA species in cells is determined by the balance between ...
Measurement of Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) in Culture Cells for Assessment of the Energy Metabolism
作者:Birte Plitzko and Sandra Loesgen日期:05/20/2018,浏览量:56327,Q&A: 0
Mammalian cells generate ATP by mitochondrial (oxidative phosphorylation) and non-mitochondrial (glycolysis) metabolism. Cancer cells are known to reprogram their metabolism using different ...
Microbial Mutagenicity Assay: Ames Test
作者:Urvashi Vijay, Sonal Gupta, Priyanka Mathur, Prashanth Suravajhala and Pradeep Bhatnagar日期:03/20/2018,浏览量:25968,Q&A: 2
The Microbial mutagenicity Ames test is a bacterial bioassay accomplished in vitro to evaluate the mutagenicity of various environmental carcinogens and toxins. While Ames test is used to ...
Stereotaxic Adeno-associated Virus Injection and Cannula Implantation in the Dorsal Raphe Nucleus of Mice
作者:Patrícia A. Correia, Sara Matias and Zachary F. Mainen日期:09/20/2017,浏览量:13476,Q&A: 1
Optogenetic methods are now widespread in neuroscience research. Here we present a detailed surgical procedure to inject adeno-associated viruses and implant optic fiber cannulas in the dorsal raphe ...
Superoxide Dismutase (SOD) and Catalase (CAT) Activity Assay Protocols for Caenorhabditis elegans
作者:Jing Zhang, Rui Chen, Zhenyang Yu and Lili Xue日期:08/20/2017,浏览量:30098,Q&A: 0
Assays for superoxide dismutase (SOD) and catalase (CAT) activities are widely employed to indicate antioxidant responses underlying the toxic effects of test chemicals. Yet, earlier studies mainly ...
Extraction of Soluble and Insoluble Protein Fractions from Mouse Brains and Spinal Cords
作者:Oliver Wirths日期:08/05/2017,浏览量:21282,Q&A: 1
The current protocol details the preparation of soluble and insoluble protein lysates from mouse brain or spinal cord samples. In detail, tissue homogenization and sequential protein extraction are ...
Measurement of the Intracellular Calcium Concentration with Fura-2 AM Using a Fluorescence Plate Reader
使用荧光读板仪通过钙指示剂Fura-2 AM 测定细胞内钙浓度
作者:Magdiel Martínez, Namyr A. Martínez and Walter I. Silva日期:07/20/2017,浏览量:25793,Q&A: 1
Intracellular calcium elevation triggers a wide range of cellular responses. Calcium responses can be affected or modulated by membrane receptors mutations, localization, exposure to ...
BODIPY 493/503 Staining of Neutral Lipid Droplets for Microscopy and Quantification by Flow Cytometry
显微镜观察BODIPY 493/503 染色中性脂滴以及用流式细胞仪量化
作者:Bo Qiu and M. Celeste Simon日期:09/05/2016,浏览量:45670,Q&A: 1
Lipid droplets (LDs) are ubiquitous, dynamic organelles and function as a storage depot for neutral lipids, including triglycerides and cholesterol esters (Walther and Farese, 2012). The movement of ...
Minimal Inhibitory Concentration (MIC) Assay for Acinetobacter baumannii
作者:Ming-Feng Lin, Yun-You Lin and Chung-Yu Lan日期:12/05/2014,浏览量:43314,Q&A: 3
Minimal inhibition concentration (MIC) is the lowest concentration of an antimicrobial agent that can inhibit the visible growth of a microorganism after overnight incubation. MIC determination is ...
Dimethylmethylene Blue Assay (DMMB)
作者:Vivien Jane Coulson- Thomas and Tarsis Ferreira Gesteira日期:09/20/2014,浏览量:49965,Q&A: 8
Glycosaminoglycans (GAGs) are long unbranched polysaccharides consisting of repeating disaccharide units composed of a hexosamine (glucosamine or galactosamine) and a hexuronic acid (glucuronic or ...