Document Detail


Silicon inhibition effects on the polymerase chain reaction: a real-time detection approach.
MedLine Citation:
PMID:  16345097     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In the miniaturization of biochemical analysis systems, biocompatibility of the microfabricated material is a key feature to be considered. A clear insight into interactions between biological reagents and microchip materials will help to build more robust functional bio-microelectromechanical systems (BioMEMS). In the present work, a real-time polymerase chain reaction (PCR) assay was used to study the inhibition effects of silicon and native silicon oxide particles on Hepatitis B Virus (HBV) DNA PCR amplification. Silicon nanoparticles with different surface oxides were added into the PCR mixture to activate possible interactions between the silicon-related materials and the PCR reagents. Ratios of silicon nanoparticle surface area to PCR mixture volume (surface to volume ratio) varied from 4.7 to 235.5 mm2/microL. Using high speed centrifugation, the nanoparticles were pelleted to tube inner surfaces. Supernatant extracts were then used in subsequent PCR experiments. To test whether silicon materials participated in amplifications directly, in some cases, entire PCR mixture containing silicon nanoparticles were used in amplification. Fluorescence histories of PCR amplifications indicated that with the increase in surface to volume ratio, amplification efficiency decreased considerably, and within the studied ranges, the higher the particle surface oxidation, the stronger the silicon inhibition effects on PCR. Adsorption of Taq polymerase (not nucleic acid) on the silicon-related material surface was the primary cause of the inhibition phenomena and silicon did not participate in the amplification process directly.
Authors:
Wei Wang; Hai-Bin Wang; Zhi-Xin Li; Zeng-Yuan Guo
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of biomedical materials research. Part A     Volume:  77     ISSN:  1549-3296     ISO Abbreviation:  -     Publication Date:  2006 Apr 
Date Detail:
Created Date:  2006-02-22     Completed Date:  2006-08-10     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  101234237     Medline TA:  J Biomed Mater Res A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  28-34     Citation Subset:  IM    
Copyright Information:
(c) 2005 Wiley Periodicals, Inc.
Affiliation:
Department of Engineering Mechanics, Tsinghua University, Beijing 100084, People's Republic of China. w.wang@pku.edu.cn
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MeSH Terms
Descriptor/Qualifier:
Hepatitis B virus / metabolism
Nanotechnology
Polymerase Chain Reaction* / instrumentation,  methods
Silicon / chemistry,  metabolism*
Spectrometry, X-Ray Emission
Chemical
Reg. No./Substance:
7440-21-3/Silicon

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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