• 卷册 12, 2022
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Image Credit: BioProtoc.2751

发育生物学

Terminal Deoxynucleotidyl Transferase Mediated Production of Labeled Probes for Single-molecule FISH or RNA Capture 末端脱氧核苷酸转移酶介导的用于单分子FISH或RNA捕获的标记探针的生成 作者:Imre Gaspar, Frank Wippich and Anne Ephrussi日期:03/05/2018,浏览量:9243,Q&A: 0
Arrays of short, singly-labeled ssDNA oligonucleotides enable in situ hybridization with single molecule sensitivity and efficient transcript specific RNA capture. Here, we describe a simple, enzymatic protocol that can be carried out using basic laboratory equipment to convert arrays of PCR oligos into smFISH and RAP probesets in a ...
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免疫学

Quantification of Bacterial Attachment to Tissue Sections 组织切片细菌附着定量 作者:Batya Isaacson, Tehila Hadad, Gilad Bachrach and Ofer Mandelboim日期:03/05/2018,浏览量:6417,Q&A: 0
Here we describe a method to test bacterial adhesion to paraffin embedded tissue sections. This method allows examining binding of different bacterial strains, transfected with a fluorescent protein reporter plasmid to various tissues, to better understand different mechanisms such as colonization. This assay provides a more physiological context ...
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Murine Pancreatic Islets Transplantation under the Kidney Capsule 肾被膜下小鼠胰岛移植 作者:Tatiana Jofra, Giuseppe Galvani, Fousteri Georgia, Gregori Silvia, Nicola Gagliani and Manuela Battaglia日期:03/05/2018,浏览量:7537,Q&A: 1
Type 1 diabetes (T1D) is an autoimmune disease caused by the lack of insulin-producing pancreatic beta cells leading to systemic hyperglycemia. Pancreatic islet transplantation is a valid therapeutic approach to restore insulin loss and to promote adequate glycemic control. Pancreatic islet transplantation in mice is an optimal preclinical model ...
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Mono Sodium Urate Crystal-induced Peritonitis for in vivo Assessment of Inflammasome Activation 单钠尿酸盐晶体诱导的腹膜炎用于体内评估炎症体激活 作者:Marianne R. Spalinger and Michael Scharl日期:03/05/2018,浏览量:7079,Q&A: 0
Due to its particulate material, mono-sodium urate (MSU) crystals are potent activators of the NOD-like receptor NLRP3. Upon activation, NLRP3 induces the formation of inflammasome complexes, which lead to the production and release of mature IL-1β. Bioactive IL-1β is a potent activator of innate immune responses and promotes recruitment of ...
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Intravenous Labeling and Analysis of the Content of Thymic Perivascular Spaces 静脉标记和胸腺血管周围间隙含量分析 作者:Roland Ruscher and Kristin A. Hogquist日期:03/05/2018,浏览量:5609,Q&A: 0
Following development in the thymus, T cells are thought to exit into the periphery predominantly through perivascular spaces (PVS). This exit route is used by conventional T cells, and likely also applies to unconventional T cell subsets, such as precursors of CD8αα and TCRγδ intraepithelial lymphocytes, regulatory T cells and natural killer T ...
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微生物学

Determination of Polyhydroxybutyrate (PHB) Content in Ralstonia eutropha Using Gas Chromatography and Nile Red Staining 采用气相色谱法和尼罗红染色法测定富养罗尔斯通菌中聚羟基丁酸酯(PHB)的含量 作者:Janina R. Juengert, Stephanie Bresan and Dieter Jendrossek日期:03/05/2018,浏览量:12837,Q&A: 0
Ralstonia eutropha H16 produces and mobilizes (re-utilizes) intracellular polyhydroxybutyrate (PHB) granules during growth. This protocol describes the visualization of intracellular Nile red stained PHB granules and the quantification of PHB by gas chromatography. Our first method describes how to analyze PHB granules by fluorescence ...
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Visualization of RNA 3’ ends in Escherichia coli Using 3’ RACE Combined with Primer Extension 3'RACE联合引物延伸法检测大肠埃希氏菌RNA 3'末端 作者:Xun Wang, Heung Jin Jeon, Monford Paul Abishek N, Jin He and Heon M. Lim日期:03/05/2018,浏览量:6696,Q&A: 0
In this assay, 3’ RACE (Rapid Amplification of cDNA 3’ Ends) followed by PE (primer extension), abbreviated as 3’ RACE-PE is used to identify the mRNA 3’ ends. The following protocol describes the amplification of the mRNA 3’ ends at the galactose operon in E. coli and the corresponding visualization of the PCR products through PE. In PE, ...
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In vivo Analysis of Cyclic di-GMP Cyclase and Phosphodiesterase Activity in Escherichia coli Using a Vc2 Riboswitch-based Assay 使用基于Vc2 Riboswitch的测定法体内分析大肠杆菌中环状di-GMP环化酶和磷酸二酯酶活性 作者:Ying Liu, Hyunhee Kim and Ute Römling日期:03/05/2018,浏览量:5711,Q&A: 0
Cyclic di-guanosine monophosphate (c-di-GMP) is a ubiquitous second messenger that regulates distinct aspects of bacterial physiology. It is synthesized by diguanylate cyclases (DGCs) and hydrolyzed by phosphodiesterases (PDEs). To date, the activities of DGC and PDE are commonly assessed by phenotypic assays, mass spectrometry analysis of ...
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神经科学

Flight and Climbing Assay for Assessing Motor Functions in Drosophila 评估果蝇运动功能的飞行和攀爬分析 作者:Steffy B Manjila and Gaiti Hasan日期:03/05/2018,浏览量:8739,Q&A: 0
Motor control requires the central nervous system to integrate different sensory inputs and convey this information to the relevant central pattern generator for execution of motor function through motor neurons and muscles. Proper motor control is essential for any mobile organism to survive and interact with the external environment. For flying ...
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Barnes Maze Procedure for Spatial Learning and Memory in Mice 用于小鼠空间学习和记忆的Barnes迷宫程序 作者:Matthew W. Pitts日期:03/05/2018,浏览量:11939,Q&A: 0
The Barnes maze is a dry-land based rodent behavioral paradigm for assessing spatial learning and memory that was originally developed by its namesake, Carol Barnes. It represents a well-established alternative to the more popular Morris Water maze and offers the advantage of being free from the potentially confounding influence of swimming ...
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A Fluorescent Dye Method Suitable for Visualization of One or More Rat Whiskers 适合观察一根或多根大鼠触须的荧光染料方法 作者:Jacopo Rigosa, Alessandro Lucantonio, Giovanni Noselli, Arash Fassihi, Erik Zorzin, Fabrizio Manzino, Francesca Pulecchi and Mathew E Diamond日期:03/05/2018,浏览量:5095,Q&A: 0
Visualization and tracking of the facial whiskers is critical to many studies of rodent behavior. High-speed videography is the most robust methodology for characterizing whisker kinematics, but whisker visualization is challenging due to the low contrast of the whisker against its background. Recently, we showed that fluorescent dye(s) can be ...
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Dual-sided Voltage-sensitive Dye Imaging of Leech Ganglia 水蛭神经节的双面电压敏感染料成像 作者:Yusuke Tomina and Daniel A. Wagenaar日期:03/05/2018,浏览量:5888,Q&A: 0
In this protocol, we introduce an effective method for voltage-sensitive dye (VSD) loading and imaging of leech ganglia as used in Tomina and Wagenaar (2017). Dissection and dye loading procedures are the most critical steps toward successful whole-ganglion VSD imaging. The former entails the removal of the sheath that covers neurons in the ...
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Mouse Phrenic Nerve Hemidiaphragm Assay (MPN) 小鼠中膈神经偏侧膈分析(MPN) 作者:Giulia Zanetti, Samuele Negro, Marco Pirazzini and Paola Caccin日期:03/05/2018,浏览量:7269,Q&A: 0
The neuromuscular junction (NMJ) is the specialized synapse by which peripheral motor neurons innervate muscle fibers and control skeletal muscle contraction. The NMJ is the target of several xenobiotics, including chemicals, plant, animal and bacterial toxins, as well as of autoantibodies raised against NMJ antigens. Depending on their ...
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Construction and Cloning of Minigenes for in vivo Analysis of Potential Splice Mutations 用于体内潜在的剪接突变分析的微小基因的构建及克隆 作者:Lisa Maria Riedmayr, Sybille Böhm, Stylianos Michalakis and Elvir Becirovic日期:03/05/2018,浏览量:9456,Q&A: 0
Disease-associated mutations influencing mRNA splicing are referred to as splice mutations. The majority of splice mutations are found on exon-intron boundaries defining canonical donor and acceptor splice sites. However, mutations in the coding region (exonic mutations) can also affect mRNA splicing. Exact knowledge of the disease mechanism of ...
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植物科学

Large Scale Field Inoculation and Scoring of Maize Southern Leaf Blight and Other Maize Foliar Fungal Diseases 玉米南方叶斑病及其他叶片真菌病的大面积野外接种与评分 作者:Shannon M. Sermons and Peter J. Balint-Kurti日期:03/05/2018,浏览量:6863,Q&A: 0
Field-grown maize is inoculated with Cochliobolus heterostrophus, causal agent of southern leaf blight disease, by dropping sorghum grains infested with the fungus into the whorl of each maize plant at an early stage of growth. The initial lesions produce secondary inoculum that is dispersed by wind and rain, causing multiple cycles of ...
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Histone Deubiquitination Assay in Nicotiana benthamiana 本氏烟草中的组蛋白去泛素化分析 作者:Shujing Liu and Lars Hennig日期:03/05/2018,浏览量:4377,Q&A: 0
Histone modifications are a group of post-translational modifications on histones which can alter chromatin structure and affect gene expression. Histone ubiquitination is a histone modification found in particular on histone H2A and H2B. Histone ubiquitination can be reversed by ubiquitin-specific proteases (UBP). Here, we describe an in vivo ...
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Boron Uptake Assay in Xenopus laevis Oocytes 在非洲爪蟾卵母细胞系统中进行硼的吸收分析实验 作者:Sheliang Wang, Namiki Mitani-Ueno and Junpei Takano日期:03/05/2018,浏览量:6118,Q&A: 0
Boron (B) is essential for plant growth and taken up by plant roots as boric acid. Under B limitation, B uptake and translocation in plants are dependent on the boric acid channels located in the plasma membrane. Xenopus leavis oocyte is a reliable heterologous expression system to characterize transport activities of boric acid channels ...
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Quantification of Plant Cell Death by Electrolyte Leakage Assay 通过电解质渗漏测定法定量植物细胞死亡 作者:Noriyuki Hatsugai and Fumiaki Katagiri日期:03/05/2018,浏览量:18562,Q&A: 2
We describe a protocol to measure the electrolyte leakage from plant tissues, resulting from loss of cell membrane integrity, which is a common definition of cell death. This simple protocol is designed to measure the electrolyte leakage from a tissue sample over a time course, so that the extent of cell death in the tissue can be monitored ...
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系统生物学

Coupling Exonuclease Digestion with Selective Chemical Labeling for Base-resolution Mapping of 5-Hydroxymethylcytosine in Genomic DNA 核酸外切酶消化联合选择性化学标记碱基分辨率定位基因组中DNA中5-羟甲基胞嘧啶 作者:Aurélien A. Sérandour, Stéphane Avner and Gilles Salbert日期:03/05/2018,浏览量:5661,Q&A: 0
This protocol is designed to obtain base-resolution information on the level of 5-hydroxymethylcytosine (5hmC) in CpGs without the need for bisulfite modification. It relies on (i) the capture of hydroxymethylated sequences by a procedure known as ‘selective chemical labeling’ (see Szulwach et al., 2012) and (ii) the digestion of the ...
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Synthetic Genetic Interaction (CRISPR-SGI) Profiling in Caenorhabditis elegans 秀丽隐杆线虫中合成遗传相互作用(CRISPR-SGI)分析 作者:John A. Calarco and Adam D. Norris日期:03/05/2018,浏览量:6529,Q&A: 0
Genetic interaction screens are a powerful methodology to establish novel roles for genes and elucidate functional connections between genes. Such studies have been performed to great effect in single-cell organisms such as yeast and E. coli (Schuldiner et al., 2005; Butland et al., 2008; Costanzo et al., 2010), ...
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