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  • Title: A strategy to study gene polymorphism by direct sequence analysis of cosmid clones and amplified genomic DNA.
    Author: Levedakou EN, Landegren U, Hood LE.
    Journal: Biotechniques; 1989 May; 7(5):438-42. PubMed ID: 2534468.
    Abstract:
    A two-step strategy is described here to rapidly analyze gene-sequence variation or polymorphism. First, DNA sequences flanking the coding region of the gene to be analyzed are determined directly from a cosmid clone, including the gene, using the modified T7 DNA polymerase and sequencing primers based on the cDNA sequence of the gene. Second, the identified gene-flanking sequences are used to design amplification primers for the polymerase chain reaction (PCR) to permit amplification of DNA segments of up to 1 kilobase in genomic DNA from multiple individuals. These amplified DNA segments are directly sequenced using the thermostable Taq DNA polymerase.
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