Next Generation Sequencing

DNA/RNA shearing

In recent years, advances in Next-Generation Sequencing (NGS) have revolutionized genomics and biology. This growth has fueled demands on upstream techniques for optimal sample preparation and genomic library construction. One of the most critical aspects of optimal library preparation is the quality of the DNA to be sequenced. The DNA must first be effectively and consistently sheared into the appropriate fragment size (depending on the sequencing platform) to enable sensitive and reliable NGS results. The Bioruptor® Pico and the Megaruptor® provide superior sample yields, fragment size, and consistency, which are essential for Next-Generation Sequencing workflows. Follow our guidelines and find the good parameters for your expected DNA size: DNA shearing with the Bioruptor®.

Programmable DNA size distribution and high reproducibility with Bioruptor® Pico using 0.65 (panel A) or 0.1 ml (panel B) microtubes. Panel A: 200 bp after 13 cycles (13 sec ON/OFF) using 100 µl volume. Average size: 204; CV%:1.89%). Panel B: 200 bp after 20 cycles (30 sec ON/OFF) using 10 µl volume. (Average size: 215 bp; CV%: 6.6%). Panel A & B: peak electropherogram view. Panel C & D: virtual gel view.

Reproducible and narrow DNA size distribution with Megaruptor® using short fragment size Hydropores Validation using two different DNA sources and two different methods of analysis. A: Shearing of lambda phage genomic DNA (20 ng/μl; 150 μl/sample) sheared at different speed settings and analyzed on 1% agarose gel. B: Bioanalyzer profiles of human genomic DNA (20 ng/μl; 150 μl/sample) sheared at different software settings of 2 and 5 kb. Three independent experiments were run for each setting. (Agilent DNA 12000 kit was used for separation and fragment sizing).

Demonstrated shearing to fragment sizes between 15 kb and 75 kb with Megaruptor® using long fragment size Hydropores.  Image shows DNA size distribution of human genomic DNA sheared with long fragment Hydropores. DNA was analyzed by pulsed field gel electrophoresis (PFGE) in 1% agarose gel and a mean size of smears was estimated using Image Lab 4.1 software.
* indicates unsheared DNA

Recommended products for DNA/RNA shearing

  • Bioruptor Pico
    Bioruptor® Pico sonication device
  • Bioruptor One
    Diagenode One sonication device
  • Megaruptor 3
    Megaruptor® 3
  • some alt
    0.1 ml Bioruptor® Pico Microtubes
  • some alt
    0.1 ml tube holder & tube adaptors for Biorupto...
  • some alt
    0.5 ml Bioruptor® Plus Microtubes
  • Product image
    0.5/0.65 ml tube holder for Bioruptor® Stan...
  • some alt
    0.65 ml Bioruptor® Pico Microtubes
  • some alt
    1.5 ml Bioruptor® Plus TPX microtubes
  • some alt
    1.5 ml Bioruptor® Plus TPX microtubes
  • some alt
    1.5 ml Bioruptor® Pico Microtubes with Caps
  • Bioruptor One - 20 ul chip
    20 μl Microfluidic Chips for the One
  • Bioruptor One - 50 ul chip
    50 μl Microfluidic Chips for the One
  • Bioruptor pico next gen sequencing
    Bioruptor® Pico sonication device
  • some alt
    Hydro Tubes
  • some alt
    Hydropore - long
  • some alt
    Hydropore - short
  • megaruptor 3 shearing kit
    Megaruptor 3 Shearing Kit
  • megaruptor
  •  DNA Shearing, RNA shearing and Chromatin shearing
    Megaruptor® 2
  • Bioruptor Pico Tube Holder
    Tube holder for 0.2 ml tubes - Bioruptor® Pico
  • Bioruptor Pico Tube Holder
    Tube holder for 0.65 ml tubes - Bioruptor® ...
  • Bioruptor Pico Tube Holder
    Tube holder for 1.5 ml tubes - Bioruptor® Pico

Related protocols

DNA Shearing with Bioruptor® Pico (2013-2019) - Small Volume (10 µl)
For DNA shearing we highly recommend to use the 0.1 ml tube holder & tube adaptors (Cat. No. ...
DNA Shearing for Bioruptor® Pico (2013-2019) - Standard 100 μl Volume
Standard protocols for programmable DNA size distributions, excellent reproducibility, and high d...
RNA Shearing for Bioruptor® NGS and Pico
The Diagenode ACT (Adaptative Cavitation Transfer technology) process is highly reproducible, how...
DNA shearing for Next-Generation Sequencing with the Bioruptor® Pico (2013-2019), 0.2 ml Bioruptor® microtubes and the corresponding holder
Optimal data generation using NGS platforms relies on a few sample-preparation prerequisites, amo...
FFPE DNA extraction protocol using the Bioruptor®
The number of archival formalin-fixed paraffin embedded (FFPE) samples is in the millions, provid...
Workflow for FFPE-derived DNA shearing prior to Next Generation Sequencing
Formalin fixation and paraffin embedding (FFPE) is a standard method for long-term preservation o...


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