Diagenode

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).


It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.

Chromatin Shearing Kit Low SDS (for Histone) Chromatin Shearing Kit Low SDS (for TF) Chromatin Shearing Kit High SDS Chromatin Shearing Kit (for Plant)

SDS concentration

< 0.1%

0.2%

1%

0.5%

Nuclei isolation

Yes

Yes

No

Yes

Allows for shearing of... cells/tissue

100 million cells

100 million cells

100 million cells

up to 25 g of tissue

Corresponding to shearing buffers from

iDeal ChIP-seq kit for Histones

iDeal ChIP-seq Kit for Transcription Factors

iDeal ChIP qPCR kit

True MicroChIP kit

Universal Plant ChIP-seq kit

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

Recommended products for Chromatin shearing

  • Bioruptor Pico
    B01080010
    Bioruptor® Pico sonication device
  • Bioruptor Plus Sonication Device
    B01020001
    Bioruptor® Plus sonication device
  • default alt
    C01019027
    ChIP Cross-link Gold
  • chip kit icon
    C01020013
    Chromatin EasyShear Kit - Low SDS
  • default alt
    C01020014
    Chromatin EasyShear Kit for Plant
  • chip kit icon
    C01020010
    Chromatin EasyShear Kit – Ultra Low SDS
  • Bioruptor One
    B01070001
    Diagenode One sonication device
  • 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
    C30010010-300
    1.5 ml Bioruptor® Plus TPX microtubes
  • some alt
    C30010010-50
    1.5 ml Bioruptor® Plus TPX microtubes
  • some alt
    C30010016
    1.5 ml Bioruptor® Pico Microtubes with Caps
  • 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 &...
  • Bioruptor pico next gen sequencing
    B01060010
    Bioruptor® Pico sonication device
  • ChIP kit icon
    C01020012
    Chromatin EasyShear Kit - High SDS
  • Bioruptor Pico Tube Holder
    B01201144
    Tube holder for 0.2 ml tubes - Bioruptor® Pico
  • Bioruptor Pico Tube Holder
    B01201143
    Tube holder for 0.65 ml tubes - Bioruptor® ...
  • Bioruptor Pico Tube Holder
    B01201140
    Tube holder for 1.5 ml tubes - Bioruptor® Pico

Related protocols

Chromatin shearing - Guide for successful chromatin preparation using the Bioruptor® Pico
Chromatin immunoprecipitation (ChIP) is a common tool used to study protein-DNA interactions in c...
Download
Chromatin shearing from tissue protocol using Diagenode’s Chromatin shearing optimization kit - Low SDS and Bioruptor® Standard, Plus or Pico
This protocol describes the chromatin preparation from fresh or frozen tissues. The isolated chro...
Download
ChIP - The Ultimate Guide for Chromatin Shearing Optimization with Bioruptor® Standard and Plus
A review of the critical points for chromatin shearing.
Download

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