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    Home > Biochemistry News > Plant Extracts News > DigestionLigationAmplification (DLA): A Simple Genome Walking Method to Amplify Unknown Sequences Flanking Mutator (Mu) Transposons and Thereby Facilitate Gene Cloning

    DigestionLigationAmplification (DLA): A Simple Genome Walking Method to Amplify Unknown Sequences Flanking Mutator (Mu) Transposons and Thereby Facilitate Gene Cloning

    • Last Update: 2020-11-08
    • Source: Internet
    • Author: User
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    Digestion–ligation–amplification (DLA), a novel
    PCR
    -based genome walking method, was developed to amplify unknown sequences flanking known sequences of interest. DLA specifically overcomes the problems associated with amplifying genomic sequences flanking high copy number transposons in large genomes. Two DLA-based strategies,
    Mu
    Clone and DLA-454, were developed to isolate
    Mu
    -tagged alleles.
    Mu
    Clone allows for the amplification of
    DNA
    flanking subsets of the numerous
    Mu
    transposons in the genome using unique three-nucleotide tags at the 3′-ends of primers, simplifying the identification of flanking sequences that co-segregate with mutant phenotypes caused by
    Mu
    insertions. DLA-454, which combines DLA with 454 pyrosequencing, permits the efficient amplification and sequencing of
    Mu
    flanking regions in a high-throughput manner.
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