A strategy to study gene polymorphism by direct sequence analysis of cosmid clones and amplified genomic DNA |
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Authors: | E N Levedakou U Landegren L E Hood |
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Institution: | Division of Biology, California Institute of Technology, Pasadena 91125. |
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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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