Increased amplification efficiency of microchip-based PCR by dynamic surface passivation.
Surface passivation is critical for effective PCR using silicon-glass chips. We tested a dynamic polymer-based surface passivation method. Polyethylene glycol 8000 (PEG 8000) or polyvinylpyrrolidone 40 (PVP-40) applied at 0.75% (w/v) in the reaction mixture produced significant surface passivation effects using either native or SiO2-precoated silicon-glass chips. PCR amplification was achieved from human genomic DNA as a template as well as from human lymphocytes. The dynamic surface passivation effect of PEG 8000 remained similar under both conditions. Dynamic surface passivation offers a simple and cost-effective method to make microfabricated silicon-glass chips PCR friendly. It can also be used in combination with static passivation (silicon oxide surface layer) to further improve PCR performance using silicon-glass PCR chips.
University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
Xing Jian
XJ
Lou
Nicholas J
NJ
Panaro
Peter
P
Wilding
Paolo
P
Fortina
Larry J
LJ
Kricka
D002469:Cell Separation; D019075:Electrophoresis, Capillary; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000096:biosynthesis / Q000235:genetics; D052250:Nitric Oxide Synthase Type III; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012825:Silicon / Q000737:chemistry; D013499:Surface Properties
C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; D012825:Silicon