Amplification and direct sequence analysis of the 23S rRNA gene from thermophilic bacteria

Ashraf Ibrahim, H. Jacob Peider Hofman-Bang, Birgitte Kiær Ahring

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    We present a simplified and fast method to obtain high-quality sequences directly from PCRs without the traditional gel purification. We also report on an improved method to obtain sequence-quality PCR products from microorganisms that are difficult to lyse with no need for DNA extraction. The technique uses exonuclease I and shrimp alkaline phosphatase to degrade residual dNTPs and primers. Our technique is shown to work on both Gram-positive and Gram-negative bacteria
    Original languageEnglish
    JournalBioTechniques
    Volume30
    Issue number2
    Pages (from-to)414-420
    ISSN0736-6205
    Publication statusPublished - 2001

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