Streamlined preparation of genomic DNA in agarose plugs for pulsed-field gel electrophoresis
Author(s) -
Linda D. Hicks,
Charlotte M. van der Graa,
Jacob Childress,
Emily Cook,
Karen Schmidt,
Frank Rosenzweig,
Eugene Kroll
Publication year - 2018
Publication title -
journal of biological methods
Language(s) - English
Resource type - Journals
ISSN - 2326-9901
DOI - 10.14440/jbm.2018.218
Subject(s) - pulsed field gel electrophoresis , agarose , computer science , agar gel , gel electrophoresis , chromatography , biology , chemistry , microbiology and biotechnology , genetics , genotype , gene
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.
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