iStock Institute of Experimental Botany, Centre of the Region Haná for Biotechnological and Agricultural Research, Olomouc, Czech Republic Jaroslav Doležel Contribution of flow cytogenetics to plant genome sequencing, gene cloning and understanding mitotic chromosome structure
22
Embed
Contribution of flow cytogenetics - uniba.sk · 2018-10-02 · iStock Institute of Experimental Botany, Centre of the Region Haná for Biotechnological and Agricultural Research,
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
iStock
Institute of Experimental Botany, Centre of the Region Haná
for Biotechnological and Agricultural Research, Olomouc, Czech Republic
Jaroslav Doležel
Contribution of flow cytogenetics to plant genome sequencing, gene cloning
and understanding mitotic chromosome structure
Mitoticchromosomes
Chromosome flow sorting
Genome sequencing
Gene cloning
Spatial DNA organization
Chromosomalproteins
Mitoticchromosomes
Contribution of flow cytogenetics to plant genome sequencing, gene cloning
and understanding mitotic chromosome structure
Sheath fluid
Deflection
plates
Excitation
light
Waste
Right
collector
Left
collector
Laser
Scattered
light
Fluorescence
emission
Relative fluorescence intensity
Chromosomes in
suspension
Flow karyotype
Flow
chamber
Num
ber
of
events
L
R
Flow cytogenetics
•Chromosome analysis rate :
~1000 chromosomes / sec
http://olomouc.ueb.cas.cz/Doležel et al., Crit. Rev. Plant Sci. 13: 275, 1994
Prerequisites
• Suspension
of intact mitotic
chromosomes
• Differences in
chromosome fluorescence
• Intact chromosomal DNA
(or proteins)
<Stained DNA
<Labeled DNA sequences
Chromosome analysis and sorting by flow cytometry
Sheath fluid
Deflection
plates
Waste
Right
collector
Laser
Scattered
light
Relative fluorescence intensity
Num
ber
of
events
Purified
chromosome fraction
Purified
chromosome fraction
L
R
•Chromosome analysis rate :
~1000 chromosomes / sec
•Chromosome yield :
2 – 5 x 105 chromosomes / day
Excitation
light
Left
collector
Flow karyotype
Flow
chamber
http://olomouc.ueb.cas.cz/
Chromosomes in
suspension
Waste
Fluorescence
emission
Doležel et al., Crit. Rev. Plant Sci. 13: 275, 1994
<Stained DNA
<Labeled DNA sequences
Flow cytogenetics
Chromosome analysis and sorting by flow cytometry
http://olomouc.ueb.cas.cz/
Marker-assisted selection
New breeding techniques (targeted genome modification)
The knowledge of genes and other DNA sequences is needed