Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1989 May 11;17(9):3425-33.
doi: 10.1093/nar/17.9.3425.

Construction of yeast artificial chromosome libraries with large inserts using fractionation by pulsed-field gel electrophoresis

R Anand  1 , A Villasante, C Tyler-Smith
Affiliations
Free PMC article

Construction of yeast artificial chromosome libraries with large inserts using fractionation by pulsed-field gel electrophoresis

R Anand et al. Nucleic Acids Res. .
Free PMC article

Abstract

A method for constructing yeast artificial chromosome (YAC) libraries with large insert sizes is reported. High molecular weight human DNA was partially digested with EcoRI and cloned in the vector pYAC4. When unfractionated DNA was used, the mean YAC size was 120kb. Fractionation by pulsed-field gel electrophoresis using a 'waltzer' apparatus to remove small DNA fragments increased the mean YAC size to congruent to 220kb or congruent to 370kb depending on the fractionation conditions. Ligated DNA prepared by this method was stable at 4 degrees C and routinely yielded transformation efficiencies of greater than 700 colonies/micrograms. It should be possible to extend the method to produce even larger inserts and to use high molecular weight DNA from any source.

PubMed Disclaimer

References

    1. Cell. 1984 May;37(1):67-75 - PubMed
    1. Science. 1987 May 15;236(4803):806-12 - PubMed
    1. Cell. 1987 Jul 31;50(3):509-17 - PubMed
    1. Nucleic Acids Res. 1987 Aug 11;15(15):5925-43 - PubMed
    1. Nature. 1988 Sep 8;335(6186):184-6 - PubMed

Publication types