diagenode Bioruptor User manual

USER MANUAL
Version 1.1
Bioruptor® Plus
Cat. No. UCD-300
Bioruptor®
Sonication System

Guarantee
Limited one year global warranty
Diagenode guarantees all products from any manufacturing defects as we rigorously test all
products to meet strict quality standards. Diagenode warrants that all standard components of
its instruments will be free of defects in materials and workmanship for a period of one (1) year
from the date that the warranty period begins, unless the original quotation or accompanying
documentation states a different warranty period. All warranty periods begin on the date of delivery
and apply only to the first purchaser of the product. If a manufacturing defect arises and a valid
claim is received within the warranty period, Diagenode, at its discretion, will repair or replace the
product in accordance with the warranty terms and conditions stated herein. In case of repair or
replacement of a product under warranty, Diagenode will cover the expenses to return the repaired
or replacement product.
This warranty covers only manufacturing defects and does not cover any damage caused by misuse,
lack of compliance to recommendations stated in the manual, neglect, accidents, abrasion, or
exposure to extreme temperatures, chemical solvents, or acids. We strongly recommend that
maintenance or repairs of Diagenode’s products are performed by our approved Diagenode service
center. Improper or incorrectly performed maintenance or repairs will void the warranty.
Technical Assistance & Ordering Information
Diagenode s.a. BELGIUM | EUROPE
Avenue de l’hôpital,1
Tour GIGA, 3rd Floor
4000 Liège - Belgium
Tel: [phone]
Fax: [phone]
[e-mail]
[e-mail]
Diagenode Inc. USA | NORTH AMERICA
400 Morris Avenue, Suite #101
Denville, NJ 07834 - USA
Tel: [phone]
Fax: [phone]
[e-mail]
[e-mail]
For a complete listing of Diagenode’s international distributors, visit:
http://www.diagenode.com/en/support/distributors.php
For the rest of the world, please contact Diagenode s.a.

PAGE 3
DIAGENODE BIORUPTOR® PLUS USER MANUAL
Diagenode Inc. North America / Phone: [phone] // Fax: [phone] // Mail: [e-mail]
Contents
Critical Steps for Maintenance and Efficient Shearing .................................... 4
Introduction ......................................................................... 5
Bioruptor® Technical Specifications .................................................... 6
Getting to Know Your Bioruptor® System ................................................ 7
Bioruptor® Components Overview ...................................................7
Sonication bath ...................................................................7
Motorized lid .....................................................................8
Metallic soundproof box. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Power Supply Adapter ............................................................. 9
Tube holders .....................................................................9
Equipment Installation .............................................................. 10
Installation overview .............................................................10
Installing the Bioruptor® Plus ......................................................12
Controlling the Sonication. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Tube Holders & Tubes ............................................................... 15
Standard protocols .................................................................. 16
DNA Shearing ...................................................................16
Chromatin Shearing ..............................................................19
Bacteria Cell Disruption ..........................................................21
Troubleshooting .................................................................... 23
Ordering Information ........................................................ Back Cover

PAGE 4 DIAGENODE BIORUPTOR® PLUS USER MANUAL
Europe Diagenode sa / CHU - Tour GIGA - B34 - 3e étage // Avenue de l’Hôpital, 1 // 4000 Liège (Sart Tilman) // Belgium // Phone: [phone] // Mail: [e-mail]
Critical Steps for Maintenance and
Efficient Shearing
General warnings
• DO NOT turn on the instrument without water.
• DO NOT tilt the sonication bath. To change the water, use either the plastic pump or a
beaker.
Water level (sonication bath)
• The sonication bath must be filled with distilled water only to the fill line (do not use deionized water). Fill line
replacement stickers can be obtained by contacting Diagenode. Change water at least once per week.
Water temperature (sonication bath)
• Optimal water temperature for sonication is 4°C. Sample temperature should not exceed 10°C.
• Methods to maintain the temperature.
- Ice - add small amounts of crushed ice (no more than 0.5 cm) to the bath every 10 minutes.
-
Water Cooler (Cat. No. BioAcc-Cool) - the flow rate of the circulating water cooler cannot exceed 500 ml per
minute for optimal sonication.
Magnetic Ultrasound Emitter Maintenance
• The ultrasound waves are created from a series of magnets that are attached to the water tank. This system is very
sensitive to the heat generated during a run.
• Do not exceed 1 hour of total sonication per run. It is critical that the machine is allowed to cool at least 20 minutes
between runs. Damage resulting from non compliance to manual instructions will void the warranty and shorten the
lifespan of the machine.
• Ultrasound Emitters can be damaged by tilting or jarring the machine. Exercise care if moving water tank.
Validated tubes for the Bioruptor® Plus
• DNA shearing: 0.5 ml Bioruptor® Microtubes for DNA Shearing (Cat. No. WA-004-0500).
• Chromatin Shearing: 1.5 ml TPX microtubes (Cat. No. M-50050 or M-50001) and 15 ml TPX tubes (Cat. No. M-UN-15).
Others tubes might be used but will require additional optimization. Once a brand of tube is optimized, switching brands
may result in changes in sonication efficiency.
Fitting 0.5 ml or 1.5 ml tubes in the corresponding tube holder
1. Place the tubes on the corresponding tube holder (0.5/0.65 ml tube holder - Cat. No. UCD-pack 0.5 or 1.5 ml tube
holder - Cat. No. UCD-pack1.5). Never leave empty spaces in the tube holder. Fill the empty spaces with tubes
containing the same volume of water. Screw the lid on the tube holder without over-tightening the lid.
2. Carefully place the tube holder on the holding plate.
3. During sonication, samples must remain at the bottom of the tube. If needed, briefly centrifuge samples during
sonication after pausing the run.
Fitting of 15 or 50 ml tubes in the corresponding tube holder
1. Loosen both the blue and the black top prior to placing the metallic reflecting bar in the tube.
2. First tighten the blue ring then the black top. This will ensure the O-ring is properly placed in the tube.
Bioruptor

PAGE 5
DIAGENODE BIORUPTOR® PLUS USER MANUAL
Diagenode Inc. North America / Phone: [phone] // Fax: [phone] // Mail: [e-mail]
Introduction
Diagenode’s Bioruptor® Plus uses a gentle method of sonication to retain
the integrity of DNA and/or biological complexes, including chromatin,
protein-protein binding, protein-DNA complexes and other biochemical
and biological assay systems. The Bioruptor® Sonication System uses
a sonication bath to generate indirect sonication waves, which emanate
from an ultrasound element below the water tank. Because the system
is gentler than other sonicators, the Bioruptor® produces better and
more consistent results than with harsher sonication methods. Up to
12 closed tubes can be sonicated in parallel and the continuous rotation
of tubes allows even distribution of the energy for efficient sonication.
The Bioruptor® enables automation of sonication, guaranteeing higher
reproducibility of results.
The effect of ultrasound on biological samples.
The Bioruptor® sonication system uses ultrasound to create focused
mechanical stress to lyse cells or shear DNA or chromatin. Ultrasound
waves pass through the sample expanding and contracting the liquid.
During expansion, negative pressures pull the molecules away from
one another to form a cavity or bubble. This process is called cavitation.
The bubble continues to absorb energy until it can no longer sustain
itself and implodes. This produces intense focused shearing forces,
that disperse and break biomolecules. The fragmentation of DNA takes
place as a consequence of this mechanical stress or shear.
With the Bioruptor®, the entire volume of water present in the sonication bath is exposed to ultrasound, allowing all the
samples to be efficiently sonicated (Figure 1). For larger tubes (15 ml or 50 ml), the transfer of the ultrasound to the
tubes is facilitated by a metallic bar in contact with the sample. This metallic bar is not a probe (no corrosion problems)
but “reflects” the ultrasound originated from the sonication bath and improves the sample sonication efficiency by a
patented resonance system (Figure 2).
Noise level measurements & precautions
CEE noise data is not applicable to the Bioruptor’s ultrasound emitter as no tests have been conducted on similar
devices to date. See listed noise levels (as measured in the Accredited Acoustics Laboratory) and precautions for the
Bioruptor®:
Leq = noise level in dBA = 78.4 dB
Lmax = dB Peak = 87.6 dB
1. Exposure limit value
The exposure limit value is the maximum amount of noise an employee may be exposed to on any single day (8 hours).
Exposure beyond this limit presents a high risk to the user.
LEXPOSURE, 8h = 87 dB(A)
PPEAK = 200 Pa (140 dB(C) referring to 20 μPa)
Fig 1. Propagation in 0.5 ml tubes and 1.5 ml tubes
Ultrasound
Transducer
Ultrasound
Transducer
Fig 2. Propagation in 15 ml & 50 ml tubes