Streamlined preparation of genomic DNA in agarose plugs for pulsed-field gel electrophoresis

Authors

  • Linda Hicks University of Montana
  • Charlotte M. van der Graaf Technical University Delft
  • Jacob Childress University of Montana
  • Emily Cook Georgia Institute of Technology
  • Karen Schmidt University of Montana
  • Frank Rosenzweig Georgia Institute of Technology
  • Eugene Kroll University of Montana

DOI:

https://doi.org/10.14440/jbm.2018.218

Keywords:

pulsed-field gel electrophoresis, deep-well 48-well plate, chromosomal DNA preparation, DNA plugs, karyotyping, genomic fingerprinting

Abstract

Genome analysis using pulsed-field gel electrophoresis (PFGE) has been used in applications ranging from typing bacterial strains to radiobiology to cancer research. While methods for running PFGE have been significantly improved since its invention, the method for preparing chromosomal DNA itself has remained essentially unchanged. This limits the applicability of PFGE, especially when analyses require many samples. We have streamlined sample preparation for routine applications of PFGE through the use of deep-well 48-well plates. Besides saving time, our protocol has the added advantage of reducing the volume of expensive reagents. Our improved protocol enables us to reduce throughput time and simplify the procedure, facilitating wider application of PFGE-based analyses in the laboratory.

Author Biography

Linda Hicks, University of Montana

Division of Biology

Research Associate Professor

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Published

2018-03-09

How to Cite

1.
Hicks L, van der Graaf CM, Childress J, Cook E, Schmidt K, Rosenzweig F, Kroll E. Streamlined preparation of genomic DNA in agarose plugs for pulsed-field gel electrophoresis. J Biol Methods [Internet]. 2018Mar.9 [cited 2021Dec.2];5(1):e86. Available from: https://jbmethods.org/jbm/article/view/218

Issue

Section

Protocols

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