Plant Chromosome Engineering: Methods and Protocols

0375427 - ÚEB 2012 RIV US eng M - Monography Chapter
Doležel, Jaroslav - Kubaláková, Marie - Čihalíková, Jarmila - Suchánková, Pavla - Šimková, Hana
Chromosome Analysis and Sorting Using Flow Cytometry.
Plant Chromosome Engineering: Methods and Protocols. New Jersey: Humana Press, 2011 - (Birchler, J.), s. 221-237. ISBN 978-1-61737-956-7
R&D Projects: GA ČR GA521/07/1573; GA MŠMT(CZ) LC06004
Institutional research plan: CEZ:AV0Z50380511
Keywords : Chromosome analysis * flow cytometry * root meristems
Subject RIV: EB - Genetics ; Molecular Biology

Chromosome analysis and sorting using flow cytometry (flow cytogenetics) is an attractive tool for fractionating plant genomes to small parts. The reduction of complexity greatly simplifies genetics and genomics in plant species with large genomes. However, as flow cytometry requires liquid suspensions of particles, the lack of suitable protocols for preparation of solutions of intact chromosomes delayed the application of flow cytogenetics in plants. This chapter outlines a high-yielding procedure for preparation of solutions of intact mitotic chromosomes from root tips of young seedlings, their analysis using flow cytometry and sorting. Seedlings are easy to handle, root meristems are karyological stable, and high degree of metaphase synchrony can be induced. Chromosomes isolated according to this protocol have well preserved morphology, withstand shearing forces during sorting and their DNA is intact and suitable for a range of applications.
Permanent Link: http://hdl.handle.net/11104/0208082