DNA-enrichment microfluidic chip for chromatin immunoprecipitation.
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.
Oh
Hyun Jik
HJ
Seoulin Bioscience Institute, 452-2 Seongnae-dong, Gangdong-gu, Seoul 134-030, Republic of Korea.
Park
Joong Yull
JY
Park
Sung Eun
SE
Lee
Bo Yun
BY
Park
Jong Sung
JS
Kim
Suel-Kee
SK
Yoon
Tae Joong
TJ
Lee
Sang-Hoon
SH
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
D000906:Antibodies; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D006657:Histones; D004247:DNA; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)