Application of the RLGS Method to Large-Size Genomes Using a Restriction Trapper
Open Access
- 1 January 1994
- journal article
- research article
- Published by Oxford University Press (OUP) in DNA Research
- Vol. 1 (2) , 99-102
- https://doi.org/10.1093/dnares/1.2.99
Abstract
We developed a method for producing restriction landmark genomic scanning (RLGS) profiles of large size genomes, such as those of higher plants or amphibians using a restriction trapper. Use of the conventional RLGS method is limited to genomes smaller than 3 × 10 9 bp, because the larger genomic DNAs, especially those of more than 1 × 10 10 bp, produce high background due to incorporation of radioactivity at non-specifically damaged sites. Our new method reduces the background levels by reducing genome complexity to 1/200–1/300 using a purification step to enrich DNA fragments carrying specific restriction landmarks at their ends using a restriction trapper. This step makes it possible to obtain RLGS patterns of larger genomes. Our paper describes the practical application for the RLGS method using a restriction trapper with the pine tree genome (3 × 10 10 bp/haploid genome; Pinus koraiensis Sieb. et Zucc.) as an example.Keywords
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