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:1306,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 ...
Efficient Agrobacterium-mediated Transformation of the Elite–Indica Rice Variety Komboka
Genetic transformation is crucial for both investigating gene functions and for engineering of crops to introduce new traits. Rice (Oryza sativa L.) is an important model in plant research, ...
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:3734,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 ...
Single Cell Migration Assay Using Human Breast Cancer MDA-MB-231 Cell Line
作者:David M. Gau and Partha Roy日期:04/20/2020,浏览量:2970,Q&A: 0
Cell migration is a fundamental cellular process that plays a crucial role in many physioglogical and pathological processes such as wound healing or cancer metastasis. Many assays have been ...
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:3753,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:11368,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 ...
Straight Channel Microfluidic Chips for the Study of Platelet Adhesion under Flow
作者:Alexander Dupuy, Lining Arnold Ju and Freda H Passam日期:03/20/2019,浏览量:8545,Q&A: 0
Microfluidic devices have become an integral method of cardiovascular research as they enable the study of shear force in biological processes, such as platelet function and thrombus formation. ...
Detection of mRNA by Whole Mount in situ Hybridization and DNA Extraction for Genotyping of Zebrafish Embryos
作者:Rachna Narayanan and Andrew C. Oates日期:03/20/2019,浏览量:8920,Q&A: 0
In situ hybridization is used to visualize the spatial distribution of gene transcripts in tissues and in embryos, providing important information about disease and development. Current ...
Induced Germinal Center B Cell Culture System
作者:Kei Haniuda and Daisuke Kitamura日期:02/20/2019,浏览量:8261,Q&A: 0
The germinal center (GC) is the site where B cells undergo clonal expansion, affinity-based selection, and differentiation into memory B cells or plasma cells. It has been difficult to elucidate ...
Induction and Analysis of Anti-CD40-induced Colitis in Mice
作者:Barbara Joyce-Shaikh, Daniel J. Cua and David Bauché日期:02/05/2019,浏览量:5927,Q&A: 0
Colon inflammation or colitis affects more than 1 million people worldwide. Several pre-clinical models, including chemical-induced (i.e., DSS, TNBS) or pathogen-induced (i.e., ...
Evaluation of Anticancer activity of Silver Nanoparticles on the A549 Human Lung Carcinoma Cell Lines through Alamar Blue Assay
作者:Nikita Sharma, Geeta Arya, R. Mankamna Kumari, Nidhi Gupta and Surendra Nimesh日期:01/05/2019,浏览量:8379,Q&A: 0
Silver nanoparticles have been widely studied to possess antimicrobial as well as anticancer activity, and have found its applications in various fields including pharmaceutical industry, ...
Analysis of the Mitochondrial Membrane Potential Using the Cationic JC-1 Dye as a Sensitive Fluorescent Probe
作者:Farzane Sivandzade, Aditya Bhalerao and Luca Cucullo日期:01/05/2019,浏览量:13871,Q&A: 0
In recent years, fluorescent dyes have been frequently used for monitoring mitochondrial membrane potential to evaluate mitochondrial viability and function. However, the reproducibility of the ...
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:21869,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:19294,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 ...
Cell Synchronization by Double Thymidine Block
作者:Guo Chen and Xingming Deng日期:09/05/2018,浏览量:23112,Q&A: 1
Cell synchronization is widely used in studying mechanisms involves in regulation of cell cycle progression. Through synchronization, cells at distinct cell cycle stage could be obtained. Thymidine ...
Isothermal Titration Calorimetry: A Biophysical Method to Characterize the Interaction between Label-free Biomolecules in Solution
作者:Andrea Saponaro日期:08/05/2018,浏览量:16895,Q&A: 0
This protocol can be applied to analyze the direct interaction between a soluble protein and a target ligand molecule using Isothermal Titration Calorimetry (ITC, Malvern). ITC allows the biophysical ...
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:53923,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 ...
Transmission Electron Microscopy for Analysis of Mitochondria in Mouse Skeletal Muscle
作者:Joseph D. McMillan and Michael A. Eisenback日期:05/20/2018,浏览量:12343,Q&A: 0
Skeletal muscle is the most abundant tissue in the human body and regulates a variety of functions including locomotion and whole-body metabolism. Skeletal muscle has a plethora of mitochondria, the ...
Generation of Luciferase-expressing Tumor Cell Lines
作者:Todd V. Brennan, Liwen Lin, Xiaopei Huang and Yiping Yang日期:04/20/2018,浏览量:12721,Q&A: 1
Murine tumor models have been critical to advances in our knowledge of tumor physiology and for the development of effective tumor therapies. Essential to these studies is the ability to both track ...
Detection and Analysis of Circular RNAs by RT-PCR
作者:Amaresh C Panda and Myriam Gorospe日期:03/20/2018,浏览量:13331,Q&A: 4
Gene expression in eukaryotic cells is tightly regulated at the transcriptional and posttranscriptional levels. Posttranscriptional processes, including pre-mRNA splicing, mRNA export, mRNA turnover, ...
Single-step Precision Genome Editing in Yeast Using CRISPR-Cas9
Genome modification in budding yeast has been extremely successful largely due to its highly efficient homology-directed DNA repair machinery. Several methods for modifying the yeast genome have ...
Quantification of Plant Cell Death by Electrolyte Leakage Assay
作者:Noriyuki Hatsugai and Fumiaki Katagiri日期:03/05/2018,浏览量:16080,Q&A: 1
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 ...
Adapting the Smart-seq2 Protocol for Robust Single Worm RNA-seq
Most nematodes are small worms that lack enough RNA for regular RNA-seq protocols without pooling hundred to thousand of individuals. We have adapted the Smart-seq2 protocol in order to sequence the ...
Isolation and Establishment of Mesenchymal Stem Cells from Wharton’s Jelly of Human Umbilical Cord
作者:Umesh Goyal, Chitra Jaiswal and Malancha Ta日期:02/20/2018,浏览量:12817,Q&A: 4
Mesenchymal stem cells (MSCs) are currently considered as ‘medicinal signaling cells’ and a promising resource in regard to cell-based regenerative therapy. Umbilical cord is a human term perinatal ...
Obtaining Acute Brain Slices
作者:Thomas Papouin and Philip G. Haydon日期:01/20/2018,浏览量:15764,Q&A: 1
Obtaining acute brain slices for electrophysiology or amperometric recordings has become a routine procedure in most labs in the field of neuroscience. Yet, protocols describing the step by step ...
Multiple Stepwise Gene Knockout Using CRISPR/Cas9 in Escherichia coli
作者:Enrico König, Francesca Zerbini, Ilaria Zanella, Davide Fraccascia and Guido Grandi日期:01/20/2018,浏览量:20331,Q&A: 1
With the recent implementation of the CRISPR/Cas9 technology as a standard tool for genome editing, laboratories all over the world are undergoing one of the biggest advancements in molecular biology ...
Targeted Nucleotide Substitution in Mammalian Cell by Target-AID
作者:Takayuki Arazoe, Keiji Nishida and Akihiko Kondo日期:06/05/2017,浏览量:11382,Q&A: 0
Programmable RNA-guided nucleases based on CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated protein) systems have been applied to various type of cells as ...
Chromatin Immunoprecipitation Experiments from Whole Drosophila Embryos or Larval Imaginal Discs
作者:Vincent Loubiere, Anna Delest, Bernd Schuettengruber, Anne-Marie Martinez and Giacomo Cavalli日期:06/05/2017,浏览量:9546,Q&A: 1
Chromatin Immunoprecipitation coupled either to qPCR (qChIP) or high-throughput sequencing (ChIP-Seq) has been extensively used in the last decades to identify the DNA binding sites of transcription ...
Muscle Histology Characterization Using H&E Staining and Muscle Fiber Type Classification Using Immunofluorescence Staining
作者:Chao Wang, Feng Yue and Shihuan Kuang日期:05/20/2017,浏览量:18419,Q&A: 1
Muscle function is determined by its structure and fiber type composition. Here we describe a protocol to examine muscle histology and myofiber types using hematoxylin and eosin (H&E) and ...
Murine Bronchoalveolar Lavage
作者:Fan Sun, Gutian Xiao and Zhaoxia Qu日期:05/20/2017,浏览量:19136,Q&A: 1
A basic Bronchoalveolar lavage (BAL) procedure in mouse is described here. Cells and fluids obtained from BAL can be analyzed by Hema3-staining, immunostaining, Fluorescence-activated cell sorting ...
Robust Generation of Knock-in Cell Lines Using CRISPR-Cas9 and rAAV-assisted Repair Template Delivery
作者:Giel Vandemoortele, Delphine De Sutter and Sven Eyckerman日期:04/05/2017,浏览量:17488,Q&A: 0
The programmable Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated nuclease 9 (Cas9) technology revolutionized genome editing by providing an efficient way to cut the ...
Polysome Fractionation to Analyze mRNA Distribution Profiles
作者:Amaresh C. Panda, Jennifer L. Martindale and Myriam Gorospe日期:02/05/2017,浏览量:21184,Q&A: 5
Eukaryotic cells adapt to changes in external or internal signals by precisely modulating the expression of specific gene products. The expression of protein-coding genes is controlled at the ...
Dot Blot Analysis of N6-methyladenosine RNA Modification Levels
作者:Lisha Shen, Zhe Liang and Hao Yu日期:01/05/2017,浏览量:20043,Q&A: 3
N6-methyladenosine (m6A) is the most prevalent internal modification of eukaryotic messenger RNA (mRNA). The total amount of m6A can be detected by several methods, ...
Sulforhodamine B (SRB) Assay in Cell Culture to Investigate Cell Proliferation
作者:Esteban A. Orellana and Andrea L. Kasinski日期:11/05/2016,浏览量:39285,Q&A: 0
The SRB assay has been used since its development in 1990 (Skehan et al., 1990) to inexpensively conduct various screening assays to investigate cytotoxicity in cell based studies (Vichai ...
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:44437,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 ...
Hemagglutination Inhibition (HI) Assay of Influenza Viruses with Monoclonal Antibodies
Heamagglutination is inhibited when antibodies are present because antibodies to the influenza virus will prevent attachment of the virus to red blood cells. The highest dilution of antibody that ...
Differentiation of THP1 Cells into Macrophages for Transwell Co-culture Assay with Melanoma Cells
作者:Michael Peter Smith, Helen Young, Adam Hurlstone and Claudia Wellbrock日期:11/05/2015,浏览量:36909,Q&A: 2
Understanding how immune cells such as macrophages interact with cancer cells is of increasing interest, as cancer treatments move towards combing both targeted- and immuno-therapies in new treatment ...
Analysis of Protein Stability by the Cycloheximide Chase Assay
Comparison of protein stability in eukaryotic cells has been achieved by cycloheximide, which is an inhibitor of protein biosynthesis due to its prevention in translational elongation. It is broadly ...
Infectious Focus Assays and Multiplicity of Infection (MOI) Calculations for Alpha-herpesviruses
作者:Anna Sloutskin and Ronald S. Goldstein日期:11/20/2014,浏览量:37123,Q&A: 0
Titration of viral stocks is a critical process before any experimental use of the virus. Here we describe an infectious focus assay for several alphaherpesviruses, a titration method for ...
Novel Object Recognition for Studying Memory in Mice
作者:Tzyy-Nan Huang and Yi-Ping Hsueh日期:10/05/2014,浏览量:27943,Q&A: 0
Memory tests are important indexes of the brain functions for rodents behavior assay. Many memory tasks require external forces (e.g. electric shocks) or intrinsic forces (e.g. ...
Targeted Gene Mutation in Rice Using a CRISPR-Cas9 System
采用CRISPR-Cas9 系统进行水稻基因定点突变
作者:Kabin Xie, Bastian Minkenberg and Yinong Yang日期:09/05/2014,浏览量:27944,Q&A: 0
RNA-guided genome editing (RGE) using bacterial type II cluster regularly interspaced short palindromic repeats (CRISPR)–associated nuclease (Cas) has emerged as a simple and versatile tool for ...
In vitro Protein Ubiquitination Assays
作者:Qingzhen Zhao and Qi Xie日期:10/05/2013,浏览量:39274,Q&A: 1
Ubiquitin can be added to substrate protein as a protein tag by the concerted actions of ubiquitin activating enzyme (E1), ubiquitin conjugating enzyme (E2) and ubiquitin protein ligase (E3). At the ...
Isolation of Mouse Tumor-Infiltrating Leukocytes by Percoll Gradient Centrifugation
作者:Ying Liu, Keqiang Chen, Chunyan Wang, Wanghua Gong, Teizo Yoshimura, Ji Ming Wang and Mingyong Liu日期:09/05/2013,浏览量:36329,Q&A: 4
A hallmark of cancer-associated inflammation is the infiltration of leukocytes into tumors, which is believed to be recruited by chemokines. Some infiltrating leukocytes such as macrophages often ...
ImmunoPrecipitation of Nuclear Protein with Antibody Affinity Columns
作者:Saïda Dadi, Dominique Payet-Bornet and Pierre Ferrier日期:02/05/2013,浏览量:30891,Q&A: 1
Co-Immunoprecipitation (Co-IP) is the method used to pull down protein partners of a protein of interest using an antibody that specifically binds to this specific protein in order to test ...
Scratch Wound Healing Assay
作者:Yanling Chen日期:03/05/2012,浏览量:113231,Q&A: 6
The scratch wound healing assay has been widely adapted and modified to study the effects of a variety of experimental conditions, for instance, gene knockdown or chemical exposure, on mammalian cell ...
Transwell Cell Migration Assay Using Human Breast Epithelial Cancer Cell
作者:Yanling Chen日期:02/20/2012,浏览量:66542,Q&A: 7
Transwell migration assays have been widely used for studying the motility of different types of cells including metastatic cancer cells. The assay is also useful in screens for compounds that act as ...