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  • Title: A simple method for direct automated sequencing of PCR fragments.
    Author: Tracy TE, Mulcahy LS.
    Journal: Biotechniques; 1991 Jul; 11(1):68-75. PubMed ID: 1835395.
    Abstract:
    A simple and rapid method for direct sequencing of PCR-generated fragments has been developed for use on Applied Biosystems 373A Automated DNA Sequencer utilizing the DyeDeoxy terminator chemistry. Standard PCR conditions are used to generate a DNA fragment, which is subsequently gel-purified to remove excess primers and unwanted PCR products. The sequencing reactions are carried out in a thermal cycler using the purified product as template DNA and the Dye-Deoxy terminators. The sequence of 500-bp region in the bacteriophage lambda genome and a 320-bp fragment of the human genomic erythropoietin gene were sequenced with greater than 99% accuracy using this method.
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