Molecular Biology

Immunophenotyping and Intracellular Staining of Fixed Whole Blood for Mass Cytometry (CyTOF)

Authors: Sangeeta Kowli and Holden Maecker, date: 09/05/2020, view: 810, 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 ...

Immunophenotyping and Intracellular Staining of Fixed Whole Blood for Mass Cytometry (CyTOF)

Authors: Sangeeta Kowli and Holden Maecker, date: 09/05/2020, view: 44, 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: 324, 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: 1239, 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: 25, 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: 668, 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 ...

Using Seed Chipping to Genotype Maize Kernels

Authors: Alison M. Mills, Lindy A. Allsman, Sareen Leon and Carolyn G. Rasmussen, date: 03/20/2020, view: 1166, Q&A: 0

Zea mays (maize) is an important model organism for studying monocot growth and development. Genotyping maize in the greenhouse or field can be time consuming and costly. Here, we describe a method to remove or chip a small amount of the ...

Different Methods of Soil DNA Extraction

Authors: Asmita Kamble and Harinder Singh, date: 01/20/2020, view: 2722, Q&A: 0

Soil is the major reservoir of microbial diversity. Only 1% of microbial diversity can be cultured while 99% is still not culturable. It is necessary to extract DNA from soil in order to explore the 99% microbial diversity, which will be useful to ...

A Simple Protocol for Informative Visualization of Enriched Gene Ontology Terms

Authors: Titouan Bonnot, Morgane B. Gillard and Dawn H. Nagel, date: 11/20/2019, view: 4223, Q&A: 1

In genome-scale datasets, Gene Ontology (GO) enrichment is a common analysis to highlight functions over-represented or under-represented in a subset of differentially expressed genes to elucidate the biological significance of the results. However, ...

Modifications in the Transformation Step of Commercially Available Site-directed Mutagenesis Kit Increase Its Price-performance

Authors: Atta Ahmad, David F. Jahad, Hamzah Karimi, Mary A. Farwell, Jean-Luc Scemama and Cindy Putnam-Evans, date: 08/05/2019, view: 1447, Q&A: 0

Site-directed mutagenesis helps unravel the relationship between the structure and function of a protein at the level of amino acid contributions. Many techniques have been established to carry out site-directed mutagenesis on a daily routine. ...

Molecular Biology

Chiasma Counting in Maize Male Meiocytes

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