Document Detail


Nanolitre droplet array for real time reverse transcription polymerase chain reaction.
MedLine Citation:
PMID:  21359328     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Recently, more and more effort has been put into the miniaturization of genetic tests such as quantitative PCR (qPCR), because it is no doubt a powerful tool for molecular diagnosis and quantitative biology. In this paper, we developed a low density nanolitre droplet array generated on a chemical modified silicon chip for gene quantification. Reliable and sensitive two step real time qRT-PCR assay for microRNA measurement was performed within 500 nL droplets. It has a dynamic range of six orders of magnitude, allowing for the quantification of microRNA input from 10(3) to 10(9) copies per reaction. We successfully applied the platform for quantitative measurement of mir-122 across five cultured cell lines. The minimum total RNA input was as low as 1 pg per assay, which showed great potential for gene quantification at single cell level. We envision the droplet based qPCR chip would be a universal and low-cost platform for gene quantification in ordinary biological laboratories.
Authors:
Yunxia Zhang; Ying Zhu; Bo Yao; Qun Fang
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-2-28
Journal Detail:
Title:  Lab on a chip     Volume:  -     ISSN:  1473-0189     ISO Abbreviation:  -     Publication Date:  2011 Feb 
Date Detail:
Created Date:  2011-3-1     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101128948     Medline TA:  Lab Chip     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou, 310058, China. yaobo08@zju.edu.cn fangqun@zju.edu.cn.
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