• 卷册 10, 2020
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Image Credit: BioProtoc.3482


卷册: 10 期号: 1


Whole-cell and Perforated Patch-clamp Recordings from Acutely-isolated Murine Sino-atrial Node Cells 小鼠窦房结细胞快速分离过程中全细胞和穿孔膜片钳记录 作者:Qadeer Aziz, Muriel Nobles and Andrew Tinker日期:01/05/2020,浏览量:1924,Q&A: 0
Cardiac pacemaker cells of the sino-atrial node are responsible for the initiation of the heart beat and express an array of ion channels. The patch-clamp technique is the gold standard method for investigating the function of ion channels expressed in electrically active cells. Conventional whole-cell and perforated patch-clamp techniques can be ...
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Analysis of Random Migration of Cancer Cells in 3D 癌细胞随机迁移的3D分析 作者:Sai P. Visweshwaran and Alexis Gautreau日期:01/05/2020,浏览量:1922,Q&A: 0
The ability of cancer cells to migrate through a complex three-dimensional (3D) environment is a hallmark event of cancer metastasis. Therefore, an in vitro migration assay to evaluate cancer cell migration in a 3D setting is valuable to examine cancer progression. Here, we describe such a simple migration assay in a 3D ...
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Acute Isolation of Cells from Murine Sino-atrial Node 小鼠窦房结细胞的快速分离 作者:Qadeer Aziz, Muriel Nobles and Andrew Tinker日期:01/05/2020,浏览量:1728,Q&A: 0
The cardiac conduction system allows the synchronized propagation of electrical activity through heart muscle. This is initiated by the spontaneous activity of the specialized pacemaker cells of the sino-atrial node (SAN). The SAN region underlies automaticity in mammals and therefore has a crucial role in the pathogenesis of cardiac disorders ...
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Steady-state and Flux-based Trehalose Estimation as an Indicator of Carbon Flow from Gluconeogenesis or Glycolysis 以基于稳态和通量的海藻糖测定作为糖异生或糖酵解碳流指标 作者:Ritu Gupta and Sunil Laxman日期:01/05/2020,浏览量:1661,Q&A: 0
Trehalose (and glycogen) is a major storage carbohydrate in many cells, including S. cerevisiae. Typically, trehalose (a disaccharide of glucose) is synthesized and stored through gluconeogenesis. However, trehalose can also be made directly from glucose, if glucose-6-phosphate is channeled away from glycolysis or pentose phosphate ...
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RNA Interactome Identification via RNA-BioID in Mouse Embryonic Fibroblasts 利用 RNA-BioID鉴定小鼠胚胎成纤维细胞中RNA相互作用体 作者:Joyita Mukherjee, Mirita Franz-Wachtel, Boris Maček and Ralf-Peter Jansen日期:01/05/2020,浏览量:1754,Q&A: 0
Cytoplasmic localization of mRNAs is common to all organisms and serves the spatial expression of genes. Cis-acting RNA signals (mostly found in the mRNA’s 3'-UTR), called zipcodes recruit trans-acting RNA-binding proteins that facilitate the localization of the mRNA. UV-cross-linking or affinity purification has been applied to identify such ...
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A Microbial Bioassay for Direct Contact Assessment of Soil Toxicity Based on Oxygen Consumption of Sulfur Oxidizing Bacteria 基于硫氧化细菌耗氧的直接接触土壤毒性的微生物检定方法 作者:Ebenezer Ashun, Umair Ali Toor, Heonseop Eom and Sang-Eun Oh日期:01/05/2020,浏览量:1393,Q&A: 0
A new direct contact assessment of soil toxicity using sulfur oxidizing bacteria (SOB) is proposed for analyzing the toxicity of soils. The proposed method is based on the ability of SOB to oxidize elemental sulfur to sulfuric acid in the presence of oxygen. Since sulfate ions are produced from sulfur by SOB oxidation activity, changes in ...
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A Yeast Chromatin-enriched Fractions Purification Approach, yChEFs, from Saccharomyces cerevisiae 酿酒酵母的染色质富集组分的纯化方法—yChEFs 作者:Abel Cuevas-Bermúdez, Ana I. Garrido-Godino, Francisco Gutiérrez-Santiago and Francisco Navarro日期:01/05/2020,浏览量:1399,Q&A: 0
We have adapted a previous procedure and improved an approach that we named yChEFs (yeast Chromatin Enriched Fractions) for purifying chromatin fractions. This methodology allows the easy, reproducible and scalable recovery of proteins associated with chromatin. By using yChEFs, we bypass subcellular fractionation requirements involved when using ...
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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:2607,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 there is a growing need for unbiased and tailored CRISPR/Cas gRNA libraries. Conventional methods for gRNA library generation apply PCR and cloning ...
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Isolation and Characterization of Live Yeast Cells from Ancient Clay Vessels 从古代陶皿中分离鉴定活酵母细胞 作者:Tzemach Aouizerat, Aren M. Maeir, Yitzhak Paz, Yuval Gadot, Amir Szitenberg, Sivan Alkalay-Oren, Shunit Coppenhagen-Glazer, Michael Klutstein and Ronen Hazan日期:01/05/2020,浏览量:2131,Q&A: 0
Ancient fermented food has been studied mainly based on residue analysis and recipes and reconstruction attempts were performed using modern domesticated yeast. Furthermore, microorganisms which participated in fermentation were studied using ancient-DNA techniques. In a recent paper, we presented a novel approach based on the hypothesis that ...
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High-throughput Site-directed Scanning Mutagenesis Using a Two-fragment PCR Approach 利用双片段PCR法进行高通量定点扫描突变 作者:Franziska M. Heydenreich, Tamara Miljuš, Dalibor Milić and Dmitry B. Veprintsev日期:01/05/2020,浏览量:1845,Q&A: 0
Site-directed scanning mutagenesis is a useful tool applied in studying protein function and designing proteins with new properties, such as increased stability or enzymatic activity. Creating a systematic library of hundreds of site-directed mutants is still a demanding and expensive task. The established protocols for making such libraries ...
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Laser Capture Micro-dissection (LCM) of Neonatal Mouse Forebrain for RNA Isolation 新生小鼠前脑激光捕获显微切割(LCM)分离RNA 作者:Achira Roy, Mei Deng, Kimberly A. Aldinger, Ian A. GLass and Kathleen J. Millen日期:01/05/2020,浏览量:2256,Q&A: 0
Precise and reproducible isolation of desired cell types or layers from heterogeneous tissues is crucial to analyze specific gene profiles and molecular interactions in vivo. Forebrain is the core site of higher functions, like cognition and memory consolidation. It is composed of heterogeneous and distinct cell types, interconnected to ...
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The Mouse Gambling Task: Assessing Individual Decision-making Strategies in Mice 小鼠博弈任务:个体决策策略评估 作者:Elsa Pittaras, Arnaud Rabat and Sylvie Granon日期:01/05/2020,浏览量:1640,Q&A: 0
Decision-making is a complex cognitive process which consists of choosing one option among several alternatives. In humans, this process is featured in the Iowa gambling task (IGT), a decision-making task that mimics real life situations by reproducing uncertain conditions based on probabilistic rewards or penalties (see Background). Several ...
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Assessing Rough-and-tumble Play Behavior in Juvenile Rats 幼年大鼠追逐打闹行为分析 作者:Jonathan W. VanRyzin, Ashley E. Marquardt and Margaret M. McCarthy日期:01/05/2020,浏览量:1736,Q&A: 0
Play is a complex social behavior that is highly conserved across mammals. In most species, males engage in more frequent and vigorous play as juveniles than females, which reflects subtle yet impactful sex differences in brain circuitry and development. In this protocol, we describe a behavioral testing paradigm to assess social play in male and ...
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Measurement of Chloroplastic NAD Kinase Activity and Whole Tissue NAD Kinase Assay 叶绿体NAD激酶活性及全组织NAD激酶活性测定 作者:Yuuma Ishikawa, Maki Kawai-Yamada and Shin-nosuke Hashida日期:01/05/2020,浏览量:1712,Q&A: 0
Nicotinamide adenine dinucleotide phosphate (NADP) synthesis requires nicotinamide adenine dinucleotide (NAD) kinase activity, substrate NAD and ATP. The NAD kinase responds to various environmental stimuli and its activity is regulated via various regulatory pathways, such as Ca2+-dependent and redox-dependent signals. Conventional in ...
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Visualization of Actin Organization and Quantification in Fixed Arabidopsis Pollen Grains and Tubes 固定拟南芥花粉粒及花粉管中肌动蛋白组织可视化及定量分析 作者:Xiaolu Qu, Qiannan Wang, Haiyan Wang and Shanjin Huang日期:01/05/2020,浏览量:1452,Q&A: 0
Although it is widely accepted that actin plays an important role in regulating pollen germination and pollen tube growth, how actin exactly performs functions remains incompletely understood. As the function of actin is dictated by its spatial organization, it is the key to reveal how exactly actin distributes in space in pollen cells. Here we ...
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