Molecular Biology

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    A One-enzyme RT-qPCR Assay for SARS-CoV-2, and Procedures for Reagent Production
    Authors:  Sanchita Bhadra, Andre C. Maranhao, Inyup Paik and Andrew D. Ellington, date: 01/20/2021, view: 11, Q&A: 0

    Given the scale of the ongoing COVID-19 pandemic, the need for reliable, scalable testing, and the likelihood of reagent shortages, especially in resource-poor settings, we have developed an RT-qPCR assay that relies on an alternative to

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    Transcardiac Perfusion of the Mouse for Brain Tissue Dissection and Fixation
    Authors:  Jinyun Wu, Yuqi Cai, Xiaoyang Wu, Yue Ying, Yilin Tai and Miao He, date: 01/05/2021, view: 57, Q&A: 0
    Transcardiac perfusion with saline followed by 4% paraformaldehyde (PFA) is widely used to clear blood and preserve brain for immunostaining or in situ hybridization. PFA breaks into formaldehyde in solution, which cross-link protein and ...
    In vitro assessment of ivermectin resistance and gene expression profiles of P-glycoprotein genes from Haemonchus contortus (L3)
    The aim of the present protocol is to improve the methodology to identify resistance to ivermectin (IVM), one of the main anthelmintic drugs against parasitic nematodes of ruminants, using the blood-feeding nematode Haemonchus contortus as ...
    Optimize Highly Efficient Genetic Transformation Method of Spring Wheat (Triticum aestivum L.) Cultivars Using Mature Embryo
    Authors:  Dongjin Kim and Megan Hager, date: 11/20/2020, view: 308, Q&A: 0
    Genetic engineering is a powerful tool to develop desired traits in crop plants to make the crops suitable for future demand and changing environments. For the genetic engineering of wheat cultivars, the development of efficient transformation and ...
    Extracellular RNA Isolation from Biofilm Matrix of Pseudomonas aeruginosa

    Most bacteria in natural ecosystems form biofilms-a bacterial community, surrounded by a polymer matrix that consists mostly of exopolysaccharides, proteins, and nucleic acids. Extracellular RNA as a matrix component is involved in biofilm

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    Immunophenotyping and Intracellular Staining of Fixed Whole Blood for Mass Cytometry (CyTOF)
    Authors:  Sangeeta Kowli and Holden Maecker, date: 09/05/2020, view: 1536, Q&A: 0
    In this report, we present the implementation of mass cytometry for intracellular staining using fixed whole blood. In our assay described here, 250 µl of whole blood, is stimulated in vitro with PMA/ionomycin (or left unstimulated), in the ...
    A Detailed and Radioisotope-free Protocol for Electrophoretic Mobility Shift Assay (EMSA)
    Author:  Nguyen Hoai Nguyen, date: 08/20/2020, view: 688, Q&A: 0
    To comprehensively characterize the functions of a transcription factor (TF), it is required to analyze the interaction of this TF with its targeted loci. Several methods such as β-glucuronidase (GUS) or luciferase reporter, yeast one-hybrid (Y1H), ...
    Heterologous Expression and Purification of SARS-CoV2 Nucleocapsid Protein
    Authors:  Ankur Garg, Lihong Liu, David D. Ho and Leemor Joshua-Tor, date: 08/05/2020, view: 1702, Q&A: 0
    This protocol describes a step by step method for heterologous expression of SARS-CoV2 Nucleocapsid (N) protein in Escherichia coli. Moreover, this protocol includes steps to purify the N protein to high purity and homogeneity. Thus, ...
    Heterologous Expression and Purification of SARS-CoV2 Nucleocapsid Protein
    Authors:  Ankur Garg, Lihong Liu, David D. Ho and Leemor Joshua-Tor, date: 08/05/2020, view: 189, Q&A: 0
    This protocol describes a step by step method for heterologous expression of SARS-CoV2 Nucleocapsid (N) protein in Escherichia coli. Moreover, this protocol includes steps to purify the N protein to high purity and homogeneity. Thus, ...
    Barley Transformation by Particle Bombardment Using Callus
    Author:  Sung-Yong Kim, date: 06/05/2020, view: 914, Q&A: 0
    The delivery of desired construct into plant cells relies mainly on Agrobacterium-mediated transformation. However, Agrobacterium transformation is still very challenging for barley as only one genotype is amendable to ...



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