PROTOCOL

Chromatin shearing with the Bioruptor® Pico, 0.2 ml
Bioruptor® Microtubes and the corresponding holder
The first critical step of a successful ChIP experiment is the preparation of sheared chromatin with a suitable
fragment range which best represents the biological scenario being investigated. The Bioruptor ® Pico ensuresthat the chromatin is efficiently sheared to an appropriate and consistent fragment size. Bioruptor ® shearing
occurs in a temperature-controlled environment providing a high quality chromatin with preserved epitopes
suitable for ChIP-qPCR and ChIP-seq experiments.

REPRODUCIBLE CHROMATIN SHEARING USING THE 0.2 ML MICROTUBES AND THE
BIORUPTOR® PICO

Reproducible chromatin shearing using the 0.2 ml microtubes and the Bioruptor® Pico. HeLa cells were fixed with formaldehyde andchromatin was prepared accordingly to Diagenode’s TrueMicroChIP Kit (Cat. No. C01010130). Samples (50,000 cells in 50 µl) were sonicated for
8 cycles of 30” ON/30” OFF with Bioruptor® Pico. Chromatin was decrosslinked, purified and analyzed by electrophoresis on agarose gel A (1 kbladder was loaded as the size standard) or by the Fragment Analyzer (Advanced Analytical Technologies).

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Please contact us for more information

Europe Diagenode sa / LIEGE SCIENCE PARK // Rue du Bois Saint-Jean, 3 // 4102 Seraing (Ougrée) // Belgium // Phone: (+32) 4 364 20 50 // E-mail: info@diagenode.com
USA Diagenode Inc. / 400 Morris Avenue, Suite 101 // Denville, NJ 07834 // USA // Phone: (+1) 862 209-4680 // Mail: info.na@diagenode.com

PROTOCOLRECOMMENDED OPERATING CONDITIONS
Chromatin shearing using the 0.2 ml microtubes for Bioruptor ® Pico allows a simultaneous shearing of 16
samples, compatible with a volume range of 20 μl to 100 μl. The total sonication time depends on many factors
like cell type, cell density, sample volume, fixation time, concentration of detergent in the shearing buffer, etc.
Hence it is important to optimize the sonication conditions for each new experiment. As best practice, choose
the shortest sonication time (which will result in a satisfactory ChIP efficiency with the highest recover andlowest background). Avoid over-sonication, as it may lead to a drop of efficiency in ChIP experiments, especiallywhen non-histone proteins are to be evaluated by ChIP.
Sample volume: 20-100 μl.

Note: a volume range of 20-50 µl will require a shorter sonication time than a 60-100 µl sample volume.
Tubes: 0.2 ml microtubes for Bioruptor ® Pico (Cat. No. C30010020).
Tube holder: Tube Holder 0.2 ml for Bioruptor ® Pico (Cat. No. B01200044) for 16 samples.
Sonication buffer: accordingly to the experimental protocol.

Note: SDS-containing buffer (0.1-1%) sonication buffer is highly recommended for an efficient cell lysis and chromatin
shearing.
Sonication cycle: 30 sec ON/30 sec OFF.
Total sonication time: varies. An initial time-course experiment of 5-10-15 sonication cycles 30’’ ON/30’’ OFF
is recommended when starting a new ChIP project using the Bioruptor ® Pico.

Please check our chromatin shearing guide athttps://www.diagenode.com/protocols/bioruptor-pico-chromatin-preparation-guide

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Chromatin Shearing Optimization Kit – High SDS (True Micro ChIP Kit)

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Chromatin Shearing Optimization Kit – Low SDS (iDeal Kit for Histones)Cat. No. C01020010

Chromatin Shearing Optimization Kit – Low SDS (iDeal Kit for TFs)

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www.diagenode.com

Please contact us for more information

Europe Diagenode sa / LIEGE SCIENCE PARK // Rue du Bois Saint-Jean, 3 // 4102 Seraing (Ougrée) // Belgium // Phone: (+32) 4 364 20 50 // E-mail: info@diagenode.com
USA Diagenode Inc. / 400 Morris Avenue, Suite 101 // Denville, NJ 07834 // USA // Phone: (+1) 862 209-4680 // Mail: info.na@diagenode.com