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. 1996 Dec 15;24(24):5054-5.
doi: 10.1093/nar/24.24.5054.

A method for high efficiency YAC lipofection into murine embryonic stem cells

J T Lee  1 , R Jaenisch
Affiliations

A method for high efficiency YAC lipofection into murine embryonic stem cells

J T Lee et al. Nucleic Acids Res. .

Abstract

We describe a modified protocol for introducing yeast artificial chromosomes (YACs) into murine embryonic stem (ES) cells by lipofection. With a decreased DNA:cell ratio, increased concentration of condensing agents and altered culture conditions, this protocol reduces the requirement for YAC DNA to a few micrograms, improves the recovery of neomycin-resistant ES colonies and increases the yield of clones containing both flanking vector markers and insert. These modifications enable generation of sufficient 'intact' transgenic clones for biological analysis with a single experiment.

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