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Molecular Methods for Evolutionary Genetics

Volume 772 of the series Methods in Molecular Biology pp 3-12

Date:

Genome Size Determination Using Flow Cytometry of Propidium Iodide-Stained Nuclei

  • Emily E. HareAffiliated withLocus of Development, Inc.
  • , J. Spencer JohnstonAffiliated withDepartment of Entomology, Texas A&M University Email author 

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Abstract

With the rapid expansion of whole-genome sequencing and other genomic studies in nonmodel ­organisms, there is a growing demand for robust and user-friendly methods for estimating eukaryotic genome sizes across a broad range of taxa. Propidium iodide (PI) staining with flow cytometry is a powerful method for genome sizing because it is relatively fast, works with a wide variety of materials, and provides information on a very large number of nuclei. In this method, nuclei are stained with PI, which intercalates into the major groove of DNA. Unknown samples are typically costained with standard nuclei of a known genome size, and the relative fluorescence is used to calculate the genome size of the unknown.

Key words

Genome size Propidium iodide Flow cytometry C-value