ELECTROPORATION OF CONFLUENT HM-1 ES CELLS LEADS TO HIGHER AMOUNT COLONIES

Authors

  • ALENA A. BENCSIK Faculty of Animal Sciences and Biotechnologies, Timişoara, România
  • I. BENCSIK Faculty of Animal Sciences and Biotechnologies, Timişoara, România
  • JANA STĂNCULEȚ Faculty of Animal Sciences and Biotechnologies, Timişoara, România
  • M. BURA Faculty of Animal Sciences and Biotechnologies, Timişoara, România

Keywords:

HM-1, embryonic stem cells, colony number, confluent, subconfluent, selection

Abstract

Electroporation is used to induce homologous recombination in the genome of the
murine ES (embryonic stem) cells. Routinelly subconfluent ES cells are
recommended to be used in such experiments. Electroporation of immunoglobulin
specific targeting vectors with different length of homology leads to reduced number
of selected colonies. The enrichment of double selected colonies is high and thus the
amount of HM-1 ES cell colonies for the analysis is very low. Here we show that the
electroporation of confluent HM-1 ES cells leads to an increased amount of simple
and double selected colonies.

References

Magin TM, McWhir J, Melton D.W. (1992): A new mouse embryonic

stem cell line with good germ line contribution and gene targeting

frequency. Nucleic Acids Res., 20, pg. 3795-3796.

Bencsik A. Alena (2008): Cercetări privind obŃinerea şoarecilor de

laborator prin manipulări genetice.Teză de doctorat, USAMVBT

Timişoara, pg.144-178.

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Published

2023-11-01