Agilent Yeast ChIP-on-chip Analysis Protocol is a protocol published by Agilent Technologies for the research of yeast ChIP-on-chip analysis technology.
This document provides a detailed step-by-step protocol for the Agilent Yeast ChIP-on-chip analysis method, version 9.2. It covers the entire workflow from sample preparation to data extraction.
The procedure begins with cell culture and cross-linking proteins to DNA using formaldehyde. Subsequent steps include cell lysis, sonication to shear DNA, immunoprecipitation of protein-DNA complexes using magnetic beads, and purification of the resulting genomic DNA. The protocol then details how to prepare blunt-end DNA, ligate it to a linker, amplify the samples via LM-PCR, and label them with Cy3 or Cy5 fluorophores for hybridization.
The protocol specifies a final concentration of 1% formaldehyde.
The cells must be incubated for 20 minutes at room temperature on a rotating platform.
Data extraction is performed using Agilent Feature Extraction Software.