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A new DNA extraction method for high‐throughput marker analysis in a large‐genome species such as Triticum aestivum
Author(s) -
Stein N.,
Herren G.,
Keller B.
Publication year - 2001
Publication title -
plant breeding
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.583
H-Index - 71
eISSN - 1439-0523
pISSN - 0179-9541
DOI - 10.1046/j.1439-0523.2001.00615.x
Subject(s) - biology , restriction fragment length polymorphism , genome , genomic dna , polyploid , dna extraction , dna , genetics , genetic marker , polymerase chain reaction , molecular marker , computational biology , gene
Gene mapping and marker‐assisted selection in complex, polyploid genomes still relies strongly on restriction fragment length polymorphism (RFLP) analysis, as conversion of RFLP to polymerase chain reaction (PCR) markers can be very difficult. DNA extraction in amounts suitable for RFLP analysis represents the most time‐consuming and labour‐intensive step in molecular marker analysis of plant populations. In this paper, a new flexible method for plant DNA extraction is presented. It allows a high‐throughput of samples in a short time without the need for freezing or lyophilizing the plant material. The method allows the isolation of genomic DNA with a yield of 100 μg for a minimal amount of 200 mg of leaf material. This is sufficient for work with large‐genome plant species such as hexaploid wheat, where 20 μg of genomic DNA are required for a single RFLP analysis.