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DNA index, genome size, and electronic nuclear volume of vertebrates from the Miami Metro Zoo
Author(s) -
Krishan Awtar,
Dandekar Payal,
Nathan Nirmal,
Hamelik Ronald,
Miller Christine,
Shaw Jackie
Publication year - 2005
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20130
Subject(s) - propidium iodide , nuclear dna , genome size , buccal swab , flow cytometry , biology , genome , dna , microbiology and biotechnology , buccal administration , genetics , mitochondrial dna , gene , apoptosis , bioinformatics , programmed cell death
Background Flow cytometry is a rapid and reliable method for measuring nuclear DNA content and genome size. Fluorochrome binding characteristics, sample preparation and differences in DNA condensation, and availability of binding sites can cause variations in results obtained. Methods Blood samples from 82 vertebrate species were collected in 10% dimethyl sulfoxide and stained with propidium iodide/hypotonic citrate or 4,6‐diamidino‐2‐phenylindole dihydrochloride for analysis of DNA content and electronic nuclear volume (ENV). Trout red blood cells (TRBCs), human peripheral blood lymphocytes, and human buccal cavity cells were used as internal standards. Results Mean fluorescence channel (MFC) values of TRBC and buccal cavity cells used as internal standards were stable at 15 to 120 min of propidium iodide staining. TRBCs mixed with other cells especially human peripheral blood cells showed an increase in MFC. ENV and MCF values were less variable in different species of birds than in reptiles or mammals. Genome size based on use of buccal cavity cells as the internal standard showed a high degree of correlation with previous reports. Conclusions Proper selection and use of internal standards and sample preparation are essential for reliable determination of DNA content and genome size in vertebrates by flow cytometry. © 2005 Wiley‐Liss, Inc.