title: Histone modification analysis by chromatin immunoprecipitation from a low number of cells on a microfluidic platform.
abstract: Histone modifications are important epigenetic mechanisms involved in eukaryotic gene regulation. Chromatin immunoprecipitation (ChIP) assay serves as the primary technique to characterize the genomic locations associated with histone modifications. However, traditional tube-based ChIP assays rely on large numbers of cells as well as laborious and time-consuming procedures. Here we demonstrate a novel microfluidics-based native ChIP assay which dramatically reduces the required cell number and the assay time by conducting cell collection, lysis, chromatin fragmentation, immunoprecipitation, and washing on a microchip. Coupled with real-time PCR, our assay permits the analysis of histone modifications from as few as ~50 cells within 8.5 h. We envision that our method will provide a new approach for the analysis of epigenetic regulations and protein-DNA interactions in general, based on scarce cell samples such as those derived from animals and patients.
authors.affiliation: Birck Nanotechnology Center, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: Tao
authors.initials: T
authors.lastname: Geng
authors.forename: Ning
authors.initials: N
authors.lastname: Bao
authors.forename: Michael D
authors.initials: MD
authors.lastname: Litt
authors.forename: Trevor G
authors.initials: TG
authors.lastname: Glaros
authors.forename: Liwu
authors.initials: L
authors.lastname: Li
authors.forename: Chang
authors.initials: C
authors.lastname: Lu
mesh_terms: D000107:Acetylation; D000906:Antibodies / Q000276:immunology; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D044127:Epigenesis, Genetic; D006657:Histones / Q000032:analysis* / Q000276:immunology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D011499:Protein Processing, Post-Translational; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D000906:Antibodies; D006657:Histones