Molecular Methods for Evolutionary Genetics pp 3-12

Part of the Methods in Molecular Biology book series (MIMB, volume 772)

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

Protocol

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 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Locus of Development, Inc.San FranciscoUSA
  2. 2.Department of EntomologyTexas A&M UniversityCollege Station, AustinUSA

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