Diagenode

Epigenetics Chromatin Analysis

Chromatin shearing

The most important steps for a successful 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.

Panel A, 10 µl volume: Chromatin samples are sheared for 10, 20 and 30 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.1 ml Bioruptor® Microtubes (Cat. No. B01200041). Panel B, 100 µl volume: Chromatin samples are sheared for 10 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.65 ml Bioruptor® Microtubes (Cat. No. WA-005-0500). Panel C, 300 µl volume: Chromatin samples are sheared for 5, 10 and 15 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using using 1.5 ml Bioruptor microtubes (Cat. No. C30010016). Prior to de-crosslinking, samples are treated with RNase cocktail mixture at 37°C during 1 hour. The sheared chromatin is then de-crosslinked overnight and phenol/chloroform purified as described in the kit manual. 10 µl of DNA (equivalent of 500, 000 cells) are analyzed on a 2% agarose gel (MW corresponds to the 100 bp DNA molecular weight marker).
 
Diagenode recommends using validated Chromatin Shearing Optimization kits with varying SDS concentrations to ensure perfect chromatin shearing:
 

SDS concentration

< 0.1%

0.2%

0.5%

1%

Nuclei isolation

Yes

Yes

Yes

No

Allows for shearing of... cells

100 million cells

100 million cells

100 million cells

100 million cells

Corresponding to shearing buffers from

iDeal ChIP-seq kit for Histones

iDeal ChIP-seq Kit for Transcription Factors

HighCell# ChIP kit

True MicroChIP kit

LowCell# ChIP kit

Table comes from our Guide for successful chromatin preparation using the Bioruptor® Pico

Made for Chromatin shearing

  • Bioruptor-pico-next-gen-sequencing
    B01060001
    Bioruptor® Pico sonication device
  • default alt
    C01019027
    ChIP Cross-link Gold
  • default alt
    C01020014
    Chromatin Shearing Optimization Kit (Universal ...
  • some alt
    C01020010
    Chromatin shearing optimization kit - Low SDS (...
  • some alt
    C01020013
    Chromatin shearing optimization kit – Low SDS (...
  • some alt
    C30010015
    0.1 ml Bioruptor® Pico Microtubes
  • some alt
    B01200041
    0.1 ml tube holder & tube adaptors for Biorupto...
  • some alt
    C30010013
    0.5 ml Bioruptor® Plus Microtubes
  • Product image
    B01200043
    0.5/0.65 ml tube holder for Bioruptor® Stan...
  • some alt
    C30010011
    0.65 ml Bioruptor® Pico Microtubes
  • some alt
    C30010016
    1.5 ml Bioruptor® Pico Microtubes with Caps
  • some alt
    C30010010-1000
    1.5 ml Bioruptor® Plus TPX microtubes
  • some alt
    C30010010-50
    1.5 ml Bioruptor® Plus TPX microtubes
  • some alt
    B01200040
    1.5 ml tube holder for Bioruptor® Pico
  • some alt
    B01200011
    1.5 ml tube holder for Bioruptor® Standard ...
  • some alt
    B01200012
    10 ml tube holder for Bioruptor® Standard & Plus
  • some alt
    C01020031
    15 ml Bioruptor® Pico Tubes & sonication beads
  • some alt
    C30010009
    15 ml Bioruptor® Plus TPX tubes
  • some alt
    B01200016
    15 ml sonication accessories for Bioruptor®...
  • some alt
    B01200013
    15 ml tube holder for Bioruptor® Standard &...
  • some alt
    B01020001
    Bioruptor® Plus sonication device
  • default alt
    C01020030
    Buffer D, chromatin shearing buffer
  • some alt
    C01020012
    Chromatin Shearing Optimization Kit - High SDS ...
  • some alt
    C01020011
    Chromatin Shearing Optimization Kit - Medium SD...
  • Product image
    C01020020
    Shearing ChIP kit - 100
  • Product image
    C01020021
    Shearing ChIP kit - 400
  • Product image
    C01020022
    Shearing Optimization kit

Events

  • Quantitative Genomics Conference 2016 (QG16)
    University College London (UCL)
    Jun 6, 2016
  • European Worm Meeting (EWM) 2016
    Berlin
    Jun 1-Jun 3, 2016
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