Introduction of large insert DNA into mammalian cells and embryos

Curr Protoc Hum Genet. 2001 Nov:Chapter 5:Unit 5.12. doi: 10.1002/0471142905.hg0512s30.

Abstract

This unit provides a set of protocols for introducing large insert DNA into cultured mammalian cells and embryos. Two different methods, spheroplast fusion and lipofection, are described for effecting transfer of YACs or gel-purified YAC DNA into cells. Additional protocols discuss preparing and transferring BACs into cells by lipofection and into embryos by microinjection.

MeSH terms

  • Animals
  • Cells, Cultured
  • Chromosomes, Artificial, Bacterial / genetics
  • Chromosomes, Artificial, Yeast / genetics
  • DNA Primers
  • DNA, Recombinant / administration & dosage*
  • DNA, Recombinant / genetics*
  • Embryo, Mammalian / cytology
  • Gene Transfer Techniques*
  • Genetic Markers
  • Genetics, Medical
  • Humans
  • Membrane Fusion
  • Mice
  • Microinjections
  • Plasmids / genetics
  • Polymerase Chain Reaction
  • Spheroplasts / genetics

Substances

  • DNA Primers
  • DNA, Recombinant
  • Genetic Markers