Nucleic Acids Research, Vol 24, Issue 2 375-379, Copyright © 1996 by Oxford University Press
MA Shoffner, J Cheng, GE Hvichia, LJ Kricka and P Wilding
The microreaction volumes of PCR chips (a microfabricated silicon chip
bonded to a piece of flat glass to form a PCR reaction chamber) create a
relatively high surface to volume ratio that increases the significance of
the surface chemistry in the polymerase chain reaction (PCR). We
investigated several surface passivations in an attempt to identify 'PCR
friendly' surfaces and used those surfaces to obtain amplifications
comparable with those obtained in conventional PCR amplification systems
using polyethylene tubes. Surface passivations by a silanization procedure
followed by a coating of a selected protein or polynucleotide and the
deposition of a nitride or oxide layer onto the silicon surface were
investigated. Native silicon was found to be an inhibitor of PCR and
amplification in an untreated PCR chip (i.e. native slicon) had a high
failure rate. A silicon nitride (Si(3)N(4) reaction surface also resulted
in consistent inhibition of PCR. Passivating the PCR chip using a
silanizing agent followed by a polymer treatment resulted in good
amplification. However, amplification yields were inconsistent and were not
always comparable with PCR in a conventional tube. An oxidized silicon
(SiO(2) surface gave consistent amplifications comparable with reactions
performed in a conventional PCR tube.
ARTICLES
Chip PCR. I. Surface passivation of microfabricated silicon-glass chips for PCR
Department of Pathology and Laboratory Medicine, School of Medicine, University of Pennsylvania, Philadelphia, 19104, USA.
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