A Microfluidic Device with Integrated Sonication and Immunoprecipitation for Sensitive Epigenetic Assays
| Autor: | Chang Lu, Zhenning Cao |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Epigenomics Immunoprecipitation Sonication Polymerase Chain Reaction Article Analytical Chemistry Cell Line Epigenesis Genetic 03 medical and health sciences Lab-On-A-Chip Devices Humans Ultrasonics Epigenetics Methylated DNA immunoprecipitation Chemistry DNA DNA Methylation Molecular biology Cell biology Chromatin 030104 developmental biology DNA methylation Chromatin immunoprecipitation |
| Zdroj: | Analytical Chemistry |
| ISSN: | 1520-6882 0003-2700 |
| Popis: | Epigenetic studies increasingly require analysis of a small number of cells that are of one specific type and derived from patients or animals. In this report, we demonstrate a simple microfluidic device that integrates sonication and immunoprecipitation (IP) for epigenetic assays, such as chromatin immunoprecipitation (ChIP) and methylated DNA immunoprecipitation (MeDIP). By incorporating an ultrasonic transducer with a microfluidic chamber, we implemented microscale sonication for both shearing chromatin/DNA and mixing/washing of IP beads. Such integration allowed highly sensitive tests starting with 100 cross-linked cells for ChIP or 500 pg of genomic DNA for MeDIP (compared to 10(6)-10(7) cells for ChIP and 1-10 μg of DNA for MeDIP in conventional assays). The entire on-chip process of sonication and IP took only 1 h. Our tool will be useful for highly sensitive epigenetic studies based on a small quantity of sample. |
| Databáze: | OpenAIRE |
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