title: DNA-enrichment microfluidic chip for chromatin immunoprecipitation.
abstract: Chromatin immunoprecipitation (ChIP) is a powerful and widely applied technique for detecting association of individual proteins with specific genomic regions; the technique requires several complex steps and is tedious. In this paper, we develop a microbead-packed microfluidic chip which eliminates most of the laborious, time-consuming, and skill-dependent processes of the ChIP procedure. A computational fluid dynamics model was established to analyze fluidic behavior in a microbead-packed microchannel. With the use of the new chip, a ChIP procedure was performed to purify the GAPDH (glyceraldehyde 3-phosphate dehydrogenase) gene from human embryonic kidney cells (cell line 293). The ChIP capability of the microfluidic chip was evaluated and compared with that of a commercial assay kit; the precipitation performance of both methods was almost identical as shown by quantitative measurement of DNA. However, our chip offers the advantage of low resin volume, and the experimental time is greatly reduced. In addition, our method is less dependent on individual technical skill. Therefore, we expect that our microfluidic chip-based ChIP method will be widely used in DNA-, gene-, and protein-related research and will improve experimental efficiency.
authors.affiliation: Seoulin Bioscience Institute, 452-2 Seongnae-dong, Gangdong-gu, Seoul 134-030, Republic of Korea.
authors.forename: Hyun Jik
authors.initials: HJ
authors.lastname: Oh
authors.forename: Joong Yull
authors.initials: JY
authors.lastname: Park
authors.forename: Sung Eun
authors.initials: SE
authors.lastname: Park
authors.forename: Bo Yun
authors.initials: BY
authors.lastname: Lee
authors.forename: Jong Sung
authors.initials: JS
authors.lastname: Park
authors.forename: Suel-Kee
authors.initials: SK
authors.lastname: Kim
authors.forename: Tae Joong
authors.initials: TJ
authors.lastname: Yoon
authors.forename: Sang-Hoon
authors.initials: SH
authors.lastname: Lee
mesh_terms: D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D002460:Cell Line; D047369:Chromatin Immunoprecipitation / Q000295:instrumentation*; D003432:Cross-Linking Reagents / Q000737:chemistry; D004247:DNA / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000276:immunology; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000235:genetics / Q000302:isolation & purification; D006657:Histones / Q000032:analysis / Q000378:metabolism; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D008956:Models, Chemical; D013997:Time Factors
chemical_list: D000906:Antibodies; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D006657:Histones; D004247:DNA; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)