caa a22 4500 397521812 CHVBK 20180308164635.0 cr unu---uuuuu 161202e199504 xx s 000 0 eng 10.1093/jxb/46.4.451 doi (NATIONALLICENCE)oxford-10.1093/jxb/46.4.451 Determining relative abundance of specific DNA sequences in flow cytometrically sorted maize nuclei [Elektronische Daten] [A. Bashir, D.P. Biradar, A. Lane Rayburn] A wide breadth of DNA content variation has been reported among maize lines. While the extent of this variation has been well documented, few studies have focused on its cause. Some of the nuclear DNA content variation has been explained by the presence of B chromosomes or knobs. However, variation in these two structures does not account for all of the observed variation. In order to identify other fluctuating DNA sequences, a rapid and reliable method of estimating relative abundance of DNA sequences needed to be developed. The potential of flow cytometry in conjunction with spot hybridization for determining relative abundance of specific DNA sequences in maize was studied. Different numbers of G1 phase nuclei were sorted on nitrocellulose filters and non-radioactive hybridization and signal detection performed. Results from these experiments revealed a significant, positive linear correlation between the amount of target sequence and signal density using both knob (R = 0.98) and ribosomal spacer (R =0.99) DNA sequences. In addition, G1 phase nuclei of eight inbred lines differing in the amount of knob heterochromatin, were sorted on to filters and the non-radioactive hybridization and signal detection performed. A significant, positive linear correlation between C-band number and signal density (R =0.83; P = 0.0051) as well as between per cent heterochromatin and signal density (R=0.96;P = 0.0002) was observed. These results indicate the usefulness of flow cytometry for spot hybridization in determining the relative abundance of DNA sequences in the maize genome. © Oxford University Press Articles nationallicence Flow cytometry nationallicence copy number nationallicence non-isotope labelling nationallicence spot hybridization nationallicence flow sorting nationallicence Zea mays L nationallicence Bashir A. National Institute for Biotechnology and Genetic Engineering Faisalabad, Pakistan aut Biradar D.P. Department of Agronomy, University of Agricultural Sciences Dharwar 580 005, Karnataka, India aut Rayburn A. Lane Department of Agronomy, University of Illinois at Urbana-Champaign 320 Plant and Animal Biotechnology Laboratory, 1201 W. Gregory Drive, Urbana, IL 61801, USA aut Journal of Experimental Botany Oxford University Press 46/4(1995-04), 451-457 0022-0957 46:4<451 1995 46 exbotj https://doi.org/10.1093/jxb/46.4.451 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/jxb/46.4.451 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Bashir A. National Institute for Biotechnology and Genetic Engineering Faisalabad, Pakistan aut NATIONALLICENCE 700 1- Biradar D.P. Department of Agronomy, University of Agricultural Sciences Dharwar 580 005, Karnataka, India aut NATIONALLICENCE 700 1- Rayburn A. Lane Department of Agronomy, University of Illinois at Urbana-Champaign 320 Plant and Animal Biotechnology Laboratory, 1201 W. Gregory Drive, Urbana, IL 61801, USA aut NATIONALLICENCE 773 0- Journal of Experimental Botany Oxford University Press 46/4(1995-04), 451-457 0022-0957 46:4<451 1995 46 exbotj Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford