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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
    B01080010
    Bioruptor® Pico sonication device
  • Bioruptor One
    B01070001
    Diagenode One sonication device
  • Megaruptor 3
    B06010003
    Megaruptor® 3
  • 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
  • Bioruptor One - 20 ul chip
    C30150001
    20 μl Microfluidic Chips for the One
  • Bioruptor One - 50 ul chip
    C30150002
    50 μl Microfluidic Chips for the One
  • some alt
    C30010018
    Hydro Tubes
  • default alt
    E07010002
    Hydropore - long
  • default alt
    E07010001
    Hydropore - short
  • megaruptor 3 shearing kit
    E07010003
    Megaruptor 3 Shearing Kit
  • DNA Shearing, RNA shearing and Chromatin shearing
    B06010002
    Megaruptor® 2
  • 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

    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. ...    		 Download
    DNA Shearing for Bioruptor® Pico (2013-2019) - Standard 100 μl Volume
    Standard protocols for programmable DNA size distributions, excellent reproducibility, and high d...    		 Download
    RNA Shearing for Bioruptor® NGS and Pico
    The Diagenode ACT (Adaptative Cavitation Transfer technology) process is highly reproducible, how...    		 Download
    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...    		 Download
    FFPE DNA extraction protocol using the Bioruptor®
    The number of archival formalin-fixed paraffin embedded (FFPE) samples is in the millions, provid...    		 Download
    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...    		 Download