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Related Concept Videos

Hybridoma Technology01:31

Hybridoma Technology

Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation, polyethylene glycol...
Genetic Screens02:46

Genetic Screens

Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...

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Genome-wide RNAi Screening to Identify Host Factors That Modulate Oncolytic Virus Therapy
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Immunoscreening after hybrid selection and translation.

B Velan1

  • 1Israel Institute for Biological Research, Ness Ziona, Israel.

Current Protocols in Molecular Biology
|February 12, 2008
PubMed
Summary

This method screens plasmid cDNA clones by selecting specific messenger RNA (mRNA). The selected mRNA is translated to identify the desired protein, enabling clone characterization.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Identifying specific gene expression is crucial in molecular biology.
  • Traditional methods for screening cDNA libraries can be laborious and time-consuming.

Purpose of the Study:

  • To develop and describe a procedure for screening plasmid cDNA clones based on their ability to select a specific mRNA.
  • To enable the identification of clones encoding desired proteins.

Main Methods:

  • Plasmid DNA immobilized on nitrocellulose filters.
  • Hybridization with messenger RNA (mRNA).
  • Elution of selected mRNA.
  • In vitro translation of eluted mRNA into 35S-labeled protein.
  • Protein identification via immunoprecipitation and SDS-PAGE.

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Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations
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Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations

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Main Results:

  • Successful selection of specific mRNA based on plasmid DNA binding.
  • Characterization of eluted mRNA through protein translation and identification.
  • Identification of plasmid clones capable of selecting mRNA that translates into the desired protein.

Conclusions:

  • The described procedure effectively screens plasmid cDNA clones for specific mRNA selection.
  • This method facilitates the identification of clones encoding target proteins.
  • The technique is adaptable for characterizing cosmids and bacteriophage DNA.