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Chromatin shearing

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
    C30010010-300
     
    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

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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