Constructing gene-enriched plant genomic libraries using methylation filtration technology.

摘要: Full genome sequencing in higher plants is a very difficult task, because their genomes are often large and repetitive. For this reason, gene targeted partial genomic becomes realistic option. The method reported here simple approach to generate gene-enriched plant libraries called methylation filtration. This technique takes advantage of the fact that repetitive DNA heavily methylated genes hypomethylated. Then, by simply using an Escherichia coli host strain harboring wild-type modified cytosine restriction (McrBC) system, which cuts containing methylcytosine, eliminated from these libraries, while low copy (i.e., genes) recovered. To prevent cloning significant proportions organelle DNA, crude nuclear preparation must be performed prior purifying DNA. Adaptor-mediated size fractionation necessary for optimal results.