Bioruptor® in Action Video
Applications | Chromatin shearing
Chromatin immunoprecipitation (ChIP) is a common method to accurately study the regulation of gene expression at the protein level. ChIP is an ideal method to uncover how a specific protein interacts with the genome.
The steps for a successful ChIP experiment are:
- Cell fixation (cross-linking) of chromatin-bound proteins such as histones or transcription factors to DNA followed by cell lysis
- Shearing of chromatin (cross-linked DNA-protein complex) by sonication to optimal fragment sizes (up to 1000 bp; depending on the desired downstream application)
- Capturing the protein-DNA complexes using specific ChIP grade or ChIP-seq grade antibodies against the histone or transcription factor of interest
- Reversal of cross-links followed by proteinase K treatment and DNA purification
- Analysis of purified DNA to identify the genomic regions where the specific protein was located by either qPCR, hybridization to microarrays (ChIP-chip) or by high-throughput ChIP-sequencing (ChIP-seq)
The most important steps for a succesful ChIP include both cell fixation and lysis, and chromatin shearing. Researchers often overlook the critical nature of both of these steps. Eliminating inconsistencies in the shearing step, Diagenode's Bioruptor® uses state-of-the-art ultrasound ACT (Adaptive Cavitation Technology) to efficiently shear chromatin. ACT enables the highest chromatin quality for high IP efficiency and sensitivity for ChIP experiments with gentle yet highly effective shearing forces. Additionally, the Bioruptor® provides a precisely controlled temperature environment that preserves chromatin from heat degradation such that protein-DNA complexes are well-preserved for sensitive, unbiased, and accurate ChIP.
Diagenode's Bioruptor® is the instrument of choice for chromatin shearing used for a number of downstream applications such as qPCR and ChIP-seq that require optimally sheared, unbiased chromatin.
|Bioruptor® Sonicator||Competitor C Sonicator||Tip/Probe Sonicators|
|Desirable fragment sizes for sequencing||NO|
|Consistent fragment sizes from 100 bp - 1000+ bp||NO|
|Consistency between samples within runs||NO|
|100% recovery of sample||NO||NO|
|Cost of operation||Low||High||Low|
The Ultimate Guide for Chromatin Shearing Optimization with Bioruptor® | Download
Time course sonication experiment with the Bioruptor® PLUS using the buffers and protocol from Diagenode's Chromatin Shearing Optimization kit - Medium SDS
Chromatin Shearing Guide