Čeština
EnglishFlow cytometric analysis and sorting of plant chromosomes
Cápal P., Said M., Molnár I., Doležel J.
Plant cytogenetics and cytogenomics: Methods and Protocols 2672: 177-200, 2023
Keywords: Accumulation of metaphase cells; Chromosome isolation; Cytogenetic stocks; FISH; FISHIS; Flow cytometry and sorting; Hydroponic; Mitotic synchrony; Plants; Seedlings
Abstract: Flow cytometry offers a unique way of analyzing and manipulating plant chromosomes. During a rapid movement in a liquid stream, large populations can be classified in a short time according to their fluorescence and light scatter properties. Chromosomes whose optical properties differ from other chromosomes in a karyotype can be purified by flow sorting and used in a range of applications in cytogenetics, molecular biology, genomics, and proteomics. As the samples for flow cytometry must be liquid suspensions of single particles, intact chromosomes must be released from mitotic cells. This protocol describes a procedure for preparation of suspensions of mitotic metaphase chromosomes from meristem root tips and their flow cytometric analysis and sorting for various downstream applications.
DOI: 10.1007/978-1-0716-3226-0_10
Fulltext: contact IEB authors
IEB authors: Petr Cápal, Jaroslav Doležel, Istvan Molnar, Mahmoud Said
Plant cytogenetics and cytogenomics: Methods and Protocols 2672: 177-200, 2023
Keywords: Accumulation of metaphase cells; Chromosome isolation; Cytogenetic stocks; FISH; FISHIS; Flow cytometry and sorting; Hydroponic; Mitotic synchrony; Plants; Seedlings
Abstract: Flow cytometry offers a unique way of analyzing and manipulating plant chromosomes. During a rapid movement in a liquid stream, large populations can be classified in a short time according to their fluorescence and light scatter properties. Chromosomes whose optical properties differ from other chromosomes in a karyotype can be purified by flow sorting and used in a range of applications in cytogenetics, molecular biology, genomics, and proteomics. As the samples for flow cytometry must be liquid suspensions of single particles, intact chromosomes must be released from mitotic cells. This protocol describes a procedure for preparation of suspensions of mitotic metaphase chromosomes from meristem root tips and their flow cytometric analysis and sorting for various downstream applications.
DOI: 10.1007/978-1-0716-3226-0_10
Fulltext: contact IEB authors
IEB authors: Petr Cápal, Jaroslav Doležel, Istvan Molnar, Mahmoud Said