Ultrasensitive PCR and real-time detection from human genomic samples using a bidirectional flow microreactor

Philip Day, Lin Chen, Jonathan West, Pierre Alain Auroux, Andreas Manz, Philip J R Day

    Research output: Contribution to journalArticlepeer-review


    In this paper we present a reliable bidirectional flow DNA amplification microreactor for processing real-world genomic samples. This system shares the low-power thermal responsiveness of a continuous flow reactor with the low surface area to volume ratio character of stationary reactors for reducing surface inhibitory effects. Silanization with dimethyldichlorosilane in combination with dynamic surface passivation was used to enhance PCR compatibility and enable efficient amplification. For real-time fragment amplification monitoring we have implemented an epimodal fluorescent detection capability. The passivated bidirectional flow system was ultrasensitive, achieving an RNase P gene detection limit of 24 human genome copies with a reaction efficiency of 77%. This starts to rival the performance of a conventional real-time PCR instrument with a reaction efficiency of 93% and revitalizes flow-through PCR as a viable component of lab on a chip DNA analysis formats. © 2007 American Chemical Society.
    Original languageEnglish
    Pages (from-to)9185-9190
    Number of pages5
    JournalAnalytical Chemistry
    Issue number23
    Publication statusPublished - 1 Dec 2007


    • analysis
    • Base Sequence
    • Dna
    • DNA Primers
    • genetics
    • Genome
    • Genome,Human
    • Human
    • Humans
    • methods
    • Miniaturization
    • Polymerase Chain Reaction
    • real-time PCR
    • Ribonuclease P
    • Sensitivity and Specificity


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