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    Home > Biochemistry News > Plant Extracts News > Constructing Gene-Enriched Plant Genomic Libraries Using Methylation Filtration Technology

    Constructing Gene-Enriched Plant Genomic Libraries Using Methylation Filtration Technology

    • Last Update: 2021-01-13
    • Source: Internet
    • Author: User
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    Full genome sequencing in higher plants is a very difficult task, because their genomes are often very large and repetitive. For this reason, gene targeted partial genomic sequencing becomes a realistic option. The method reported here is a simple approach to generate gene-enriched plant genomic libraries called methylation filtration. This technique takes advantage of the fact that repetitive
    DNA
    is heavily methylated and genes are hypomethylated. Then, by simply using an
    Escherichia coli
    host strain harboring a wild-type modified cytosine restriction (McrBC) system, which cuts DNA containing methylcytosine, repetitive DNA is eliminated from these genomic libraries, while low copy DNA (i.e., genes) is recovered. To prevent cloning significant proportions of organelle DNA, a crude nuclear preparation must be performed prior to purifying genomic DNA. Adaptor-mediated cloning and DNA size fractionation are necessary for optimal results.
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