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Published on: February 18, 2010
Flow cytometric chromosome sorting in plants: the next generation
Jan Vrána1, Hana Simková, Marie Kubaláková
1Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic.
Methods (San Diego, Calif.)
|March 24, 2012
Summary
Flow cytometry enables chromosome purification for plant genome analysis. This method, combined with next-generation sequencing, offers a cost-effective approach to complex plant genomes.
Area of Science:
- Genomics
- Plant Biology
- Molecular Biology
Background:
- Large plant genomes and sequence redundancy from repetitive DNA and polyploidy complicate genome analysis.
- Dissecting genomes into single chromosomes is a potential solution to reduce sample complexity.
Purpose of the Study:
- To present a procedure for preparing chromosomal DNA for next-generation sequencing.
- To enable targeted and cost-effective analysis of complex plant genomes.
Main Methods:
- Utilizing flow cytometric sorting for the purification of large numbers of single chromosomes.
- Developing a DNA preparation protocol compatible with next-generation sequencing.
Main Results:
- Successful purification of chromosomes using flow cytometry.
- Generation of chromosomal DNA suitable for downstream next-generation sequencing.
Conclusions:
- Flow cytometric chromosome sorting coupled with next-generation sequencing is a powerful strategy for analyzing complex plant genomes.
- The described methods facilitate efficient and cost-effective genome dissection and sequencing.
