Rapid cloning of mammalian cells with honeycomb cloning plates and nonlethal vital stains

Robert J. Klebe

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

A rapid and technically simple method for cloning both adhesive and nonadhesive mammalian cells is described. The procedure employs (a) honeycomb cloning plates and (b) nonlethal vital stains. Instead of placing cloning rings around colonies, cells are initially seeded at clonal density directly into a plate containing an array of cloning rings (the honeycomb plate). Hence, the time involved in placing cloning rings around colonies is eliminated. Second, clone-containing wells of the honeycomb plate are easily identified by staining plates with the nonlethal vital stains, MTT or INT tetrazolium. Vital staining eliminates the time involved in searching for clones. Last, clones are transferred with a cotton-tipped swab thereby eliminating the time involved in trypsinization of cells. In this fashion, one can pick and transfer clones ofsubstrate adherent mammalian cells at a rate of one clone/ 10 to 15 s. Thus, mammalian cells can be cloned as rapidly as cloning can be carried out in microbial systems.

Original languageEnglish (US)
Pages (from-to)127-132
Number of pages6
JournalIn Vitro
Volume20
Issue number2
DOIs
StatePublished - Feb 1984

Keywords

  • cloning
  • monoclonal antibodies
  • somatic cell genetics

ASJC Scopus subject areas

  • Biotechnology
  • Plant Science

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