QIAamp Media MDx Kit (12)

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Publication protocol

Beckman Coulter Genomics received 2,150 clinical cervical samples and extracted DNA using a Qiagen QIAamp media MDx kit (Qiagen, Valencia, CA) and the Qiagen BioRobot universal system (Qiagen, Valencia, CA). Each DNA sample was PCR amplified using the β-globin and PGMY consensus HPV primers (16), and 10 μl of the amplicon was analyzed by agarose gel electrophoresis. Samples that tested positive for HPV in the PGMY PCR were reextracted in triplicate. The extractions were pooled and PCR amplified with each of 16 HPV-specific primers (the 14 hrHPV types described above, as well as HPV73 and HPV82). The amplicon size varied from 398 bp for HPV73 to 692 bp for HPV31. PCR products were purified enzymatically using exonuclease and shrimp alkaline phosphatase and sequenced on each strand using the subtype-specific forward and reverse primers to achieve bidirectional coverage.

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Manufacturer protocol

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