Microfluidic Tools for DNA Analysis - DNA Nanotechnology: From Structure to Function - page 119

Fig. 7.5 Solid-phase amplification ( SPA ). ( a ) Strict SPA. Primers (up 1 or up 2 )are5 0 -end
covalently attached to the solid support. Thermocycling the surface ( H hybridization, E elongation,
D denaturation) in the presence of a solution containing DNA template, dNTPs, and a thermostable
polymerase leads to the amplification of DNA molecules on the solid surface. During the
initial cycle, only interfacial amplification occurs, as the DNA template is present only in
solution. During subsequent cycles, in addition to interfacial amplification, surface amplification
also occurs between the copied DNA template (5 0 -end attached to the surface) and attached
primers in the vicinity (Reproduced from Ref. [ 28 ] with permission of Oxford University Press).
( b ) SPA consisting essentially of on-surface reaction and in-liquid phase reaction (Reprinted
with permission from Ref. [ 29 ]. Copyright 2009, American Institute of Physics). ( c )On-chip
multiplexed SPA. The DNA probes for matrix ( M ) gene, and hemagglutinin ( HA ) genes specific for
H5 and H7 are represented as gray circles in the upper schematic. Each dot is 150 m in diameter
with a center-to-center distance of 300 m. Green circles are guiding dots used for orientation.
Fluorescent images after 40-cycle amplification of four different viral RNA strains: AIV H1N1,
AIV H5N1, AIV H7N5, and Newcastle disease virus ( NDV ). M, primers targeting M gene which
is conserved across all type A influenza viruses; H5, primers targeting H5 regions on HA gene of
influenza virus with avian origin; H7, primers targeting H7 regions on HA gene of influenza virus
with avian origin (Reproduced from Ref. [ 30 ] with permission of The Royal Society of Chemistry)