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Evaluating a High‐throughput method of Shearing DNA Using a Bead Mill for ChIP
Author(s) -
Easparro Brandon
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.07462
Subject(s) - chromatin immunoprecipitation , chromatin , micrococcal nuclease , dna , chip , shearing (physics) , histone , nanotechnology , materials science , chromatography , chemistry , computer science , gene , composite material , promoter , biochemistry , gene expression , telecommunications , nucleosome
Chromatin Immunoprecipitation (ChIP) is key to understanding DNA‐protein interactions. This includes the mapping transcriptional factors and histone modifiers. Recent advancements have shown a transition from ChIP‐chip experiments to ChIP‐seq experiments, partly due to a reduction in materials cost, an increase in genomic coverage, as well as a reduction in required starting material. However, preparation of the chromatin is still prepared by either shearing cross‐linked chromatin by sonication or native chromatin micrococcal nuclease digestion. These options require longer incubation times and lower throughput, resulting in overall longer preparation. Bead milling provides an alternative of shearing DNA through the use of high impact beads. Through the use of specific beads, DNA shearing is achievable. Here we report the comparison of bead milling to ultrasonic shearing of DNA extracted from HEK cells for ChIP analysis. Support or Funding Information Omni International, Inc 935 Cobb Place Blvd Kennesaw Ga 30144