Bioruptor^®

Bioruptor 超声破碎仪，采用自适应空化技术，具有最高剪切性能

Diagenode Bioruptor 是最先进的超声处理系统，在超过 4,000 篇出版物中受到好评。Bioruptor 可同时处理多达 16 份样品，可灵活选择 0.1 mL 至 50 mL 的管，从而准确实现不同片段范围的随机和无偏剪切。封闭管形式可防止样品交叉污染，从而实现无与伦比的重现性。Bioruptor 使用高度受控的超声波能量及 ACT（自适应空化技术，一种非接触式、并行处理和等温技术），是用于染色质免疫沉淀 (ChIP) 的染色质剪切、用于下一代测序的基因组 DNA 剪切、RNA 剪切、细胞和组织破碎等的理想选择。

转到 Bioruptor 产品选择器表

在以下研究领域，出色的超声处理至关重要

神经科学与神经疾病

干细胞发育

癌症研究

植物和农业

环境和毒理学

Bioruptor 可满足您在所有这些领域以及更多领域的超声处理需求：

• 染色质分析
  将染色质剪切至下游 ChIP-seq 和 ChIP-qPCR 所需的精确片段大小范围。

  成功进行 ChIP 的最重要步骤包括细胞固定和裂解以及染色质剪切。Diagenode Bioruptor 使用最先进的超声技术，以温和而高效的剪切力为 ChIP 实验提供最高染色质质量，从而获得较高的 IP 效率和灵敏度。此外，Bioruptor 还提供了一种精确受控的温度环境，可防止染色质发生热降解，从而使蛋白-DNA 复合物得到良好保存以用于敏感、无偏和准确的 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).

• 临床应用
  在不使用溶剂的情况下，温和、彻底地使 FFPE 样品脱蜡

  FFPE 核酸提取

  FFPE 样品的脱蜡通常使用非极性溶剂（如二甲苯）或矿物油法进行，既费时又麻烦。Diagenode Bioruptor 是一种更优良的方法：在一个无溶剂步骤中去除石蜡并进行 FFPE 组织复水，随后进行轻度交联逆转以保持 DNA 和 RNA 的完整性。使用 Bioruptor 通过超声处理使 FFPE 切片有效脱蜡。用 Bioruptor Pico 对 10 µm 切片进行超声处理 3 个周期（室温下 30 sec 开启/关闭）。石蜡已被乳化并与组织切片完全分离。染色质剪切。成功的 ChIP 的最重要步骤包括细胞固定和裂解以及染色质剪切。Diagenode Bioruptor 使用最先进的超声技术，以温和而高效的剪切力为 ChIP 实验提供最高染色质质量，从而获得较高的 IP 效率和灵敏度。此外，Bioruptor 还提供了一种精确受控的温度环境，可防止染色质发生热降解，从而使蛋白-DNA 复合物得到良好保存以用于敏感、无偏和准确的 ChIP。

• 下一代测序：DNA、RNA 和 DNA 甲基化研究
  将 DNA 剪切至下游基因库制备所需的精确片段大小范围 -- 低进样体积低至 5 µL！

  DNA 剪切

  下一代测序 (NGS) 彻底改变了基因组学和生物学。最佳基因库制备的最关键方面之一是待测序的 DNA 的质量。首先，必须将 DNA 有效且一致地剪切成适当的片段大小（取决于测序平台）以获得敏感且可靠的 NGS 结果。Bioruptor 和 Megaruptor® 提供了出色的样品产量、片段大小和一致性，这对于下一代测序工作流程至关重要。

  

  Programmable DNA size distributions, excellent reproducibility, and high dsDNA yields with the 0.2ml tube holder for Bioruptor Pico. Figure A shows different DNA size distributions of sheared genomic DNA produced by varying the duration of sonication. The different curves depict a specific Bioruptor® Pico run, optimized to produce specific mean sizes and size ranges for NGS. Figure B-D show the excellent reproducibility in DNA shearing compatible with NGS librairies from Illumina, IonTorrent and exom capture protocols from Agilent and Roche NimbleGen requirements. All DNA samples (100ng-4ug/50ul) were analysed on Fragment Analyzer™ (Advanced Analytical).

  RNA shearing

  RNA sequencing is a highly accurate and sensitive method to obtain unprecedented information about the transcriptome. The RNA must be fragmented to an appropriate size for sequencing prior to reverse transcription. The Bioruptor provides unbiased RNA shearing for best cDNA synthesis and ensures high quality Next-Generation Sequencing.

  

  Programmable RNA size distribution and excellent reproducibility with Bioruptor. The various panels show different RNA size distributions of sheared total RNA produced by varying the duration of sonication on the Bioruptor. Panel A shows duplicate profiles produced after 5 (lanes 2-3), 10 (lanes 4-5) and 15 minutes (lanes 6-7) (30 sec on/off) of sonication. Lane 1 shows the unfragmented total RNA (starting material). Panel B and C compare the RNA size distributions of sheared total RNA from 2 different experiments. All samples were analysed on Biorad Experion using Eukaryote Total RNA HighSens chip.

• RNA 提取
  RNA 提取试剂盒与 Bioruptor 组合可维持 RNA 的完整性，并确保基因表达分析的最大产量。需要对动物组织进行有效破碎和均质化，以确保 RNA 的高产量。

  各种生化和分析技术都需要从组织中提取 RNA。分离完整 RNA 对于基因表达分析中使用的许多技术至关重要。为了获得 RNA 的最佳产量，需要对组织和培养细胞进行有效破碎和均质化。Diagenode Bioruptor Plus 使用最先进的超声 ACT（自适应空化技术）结合独特装置（如用作超声“增强剂”的微珠或管）或专用试剂（如我们的 RNA 提取试剂），只需一步，即可有效破碎组织和培养细胞，从而提供高质量 RNA 提取。

  Read more about our RNA Extraction Kit

• 质谱
  分离蛋白质用于研究或制药应用技术。

  用于显微切割的新鲜冷冻组织样品的超灵敏蛋白质组学

  本方案专为超灵敏蛋白质组学设计，能够对显微切割组织样品进行高通量定量蛋白质组学研究。Bioruptor® Pico 提供了高效的蛋白质提取，对于微型组织样品中的蛋白质分析至关重要。

  了解更多

  使用 Bioruptor 处理样品，可更有效地分离要通过质谱进行分析的蛋白质

  基于质谱 (MS) 的蛋白质组学通常需要多个样品处理步骤。复杂的工作流程、大量的样品分馏和蛋白水解非常耗时且限制了技术重现性。在本应用说明中，我们展示了简单可靠的 in-StageTip (iST) 试剂盒结合 Bioruptor Pico 样品超声处理在哪些方面优于不使用超声处理或仅使用热处理的样品制备工作流程。

  阅读本应用说明

  有关质谱和完美蛋白质分离的其他文章：
  质谱 – 使用 Bioruptor 裂解秀丽隐杆线虫用于生化和蛋白质组学分析
  质谱：鸟枪法蛋白质组学程序

• 蛋白质提取
  获得用于下游表征（如 PAGE、免疫印迹、质谱或蛋白质纯化）的高质量、高产量蛋白质

  各种生化和分析技术需要从组织或哺乳动物、酵母和细菌细胞中提取蛋白质。获得高质量、高产量蛋白质对于进一步下游蛋白质表征（如 PAGE、免疫印迹、质谱或蛋白质纯化）至关重要。使用 Diagenode Bioruptor，只需一步，即可对组织和培养细胞进行有效破碎和均质化处理，从而提供高质量蛋白质。

  

  Simultaneous extraction of cytoplasmic, nuclear and chromatic proteins
  
  Protein staining of cell lysates (containing both chromatin and soluble proteins) obtained with the protocol developed by Lauriane Fritsch from the laboratory of Slimane AIT-SI-ALI. A. Ponceau S Staining Western Blot of Hep G2 cells lysates obtained usting the Bioruptor. The rectangle indicates the location of histones. B. Western Blot of Hep G2 cells transfected and cultivated for 72h with siRNA against hMOF, a histone h4 K16-specific acetyl transferase. Both soluble (hMOf and B-actine) and chromatin (Histone 3 and 4) proteins are obtained on the same cell extract.

  

  Most reliable and reproducible method to prepare lysates from C. elegans
  
  Prasad Kasturi and F Ulrich Hartl at the Max Plank Institute for Biochemistry are using the Bioruptor for their research using C. elegans. SILAC quantitative proteomics showing the high reproducibility of proteome analysis in multiple replicates of worm samples of different ages (adopted from Walther DM and Kasturi P et al., Cell. 2015).

  Read more about our protein extraction kit

  Read more application notes for protein extraction:
  Tissue and cell extraction
  Western blot

• 蛋白质溶解
  温和轻松地溶解蛋白质，用于疾病研究。

  Neuronal proteins α-synuclein and tau are prone to pathological misfolding and aggregation in certain neurodegenerative diseases. Sonication of such proteins is conventionally used to create seeds that enable the investigation of the mechanisms that underlie the formation of misfolded protein and its propagation in cellular and animal models of disease. Sonicated fibers and seeds have a greater ability to seed protein formation from a pool of un-aggregated protein. The sonication process achieves this by breaking or shearing protein fibers into smaller pieces opening up more ends for elongation. However, standardizing input seeding material can be difficult. Conventional sonication with an immersed probe can produce variable results due to poor control of temperature and energy being passed into the sample

  The Diagenode Bioruptor® on the other hand utilizes a sonication bath-based rotor in which tubes are rotated through an ultrasound field allowing for consistent exposure of energy. In addition, the combination of the Bioruptor’s ultrasound and isothermal processing preserve protein integrity and maximize sample recovery. Read a recent application note from Dr. Kelvin Luk who demonstrated that the Bioruptor® provides a number of benefits in standardizing and creating representative seeding material that can serve to nucleate inclusions in neurons to better understand protein aggregation and misfolding models in α-synuclein in Parkison’s disease.

• 脂质体
  Bioruptor 可提供高通量和重现性，且无需直接接触，从而防止样品污染。

  超声处理是脂质体制备最常用的方法之一。Bioruptor 可提供高通量和重现性，且无需直接接触，从而防止样品污染。精确温度控制可防止脂质因过热或氧化而受损。

  

  Size reduction of multilamellar vesicle
  
  Peter Stone and Yvonne Perrie from Aston University and University of Strathclyde process MLV generated using the thin film lipid hydration method with the Bioruptor. The 100 µl samples are sonicated 15 minutes at 45°C in the Bioruptor Plus with high power for rapid small scale production of bilayer -loaded liposomes.

Bioruptor 技术的工作原理

Bioruptor Pico 和 Plus 使用基于超声浴的转子。超声浴的壁以随机但可重现的模式反射超声波。管座中的样品旋转通过超声场，以使每份样品均暴露于相同的能量水平和强度，以确保剪切一致性。独特的冷却系统，可提供等温处理和温和的超声，从而保护并保持生物样品的完整性并确保高样品回收率。

• 了解更多
  

  Figure: Isothermal and homogeneous sample preparation with the Bioruptor. Left panels: Precise temperature control to maintain integrity in sample preparation. The single cycle valve and the 4°C water cooler deliver efficient cooling and reliable performance. Finite element analysis software modeling of temperature around the sample (upper panel). Sample temperature measured with Krystal MV64 thermocouple during sonication cycles (30’’/30’’ cycles) showing the perfect isothermal processing (lower panel). Right panels: Powerful and uniform processing of samples down to 5ul (12 samples) to large volumes of 20ml (3 samples). Bioruptor empowers optimal and reproducible chromatin shearing while preserving high protein integrity (upper panel). Bioruptor’s most advanced ultrasound transmitter and water bath design ensure equal energy distribution. Cavitation energy, expressed as % of the maximum level, is measured with PPB Ultrasonic/Megasonic energy meter (lower panel).

  What are the effects of ultrasound on biological samples?

  High powered ultrasound waves can produce gaseous cavitation in liquids. Cavitation is the formation of small bubbles of dissolved gases or vapors due to the alteration of pressure in liquids. These bubbles are capable of resonance vibration and produce vigorous eddying or microstreaming. This mechanical stress has multiple effects on biological samples including; effective chromatin shearing, DNA & RNA shearing, cell lysis and tissue homogenization. When using a probe sonicator, the microstreaming phenomenon is limited to the vicinity of the probe which can generate high amounts of heat and release metal fragments. In contrast, the Bioruptor sonication bath is equally exposed to ultrasound energy allowing for the dissipation of heat and providing uniform absorption of energy. How does it work? Ultrasonic waves generated by the transducer produce longitudinal vibrations (alternating compression and rarefactions) in the water bath (see figure below). These pressure fluctuations create millions of microscopic bubbles (cavities) caused by the negative pressures generated by the rarefaction stage. As the cavities are exposed to the positive pressures, produced by the compression stage, they oscillate and expand to an unstable size (up to 100 microns diameter). Finally the cavities will implode generating pressure extremes and temperatures at the implosion sites. In addition, shock waves and eddies radiate outwardly from the site of cavity collapse. The localized hot spots that last for microseconds can have temperatures of ~5,000ºC, pressures of ~500 atmospheres as well as heating and cooling rates >109 K/s. The cumulative energy generated by this cavitation phenomenon is extremely high and produces intense shearing forces (Figure 1).

  
  Figure 1: Presentation of bubble size in function of time during sonication process.

  Bioruptor is the most popular brand in the Epigenetic and Next-Generation Sequencing communities with more than 2000 citations in peer-reviewed journals. Our instruments provide processing performance and recovery unmatched by any other sample preparation methodology. Bioruptor high intensity emission of ultrasounds creating cavitation is ideal for Epigenetics studies of chromatin and DNA methylation, Next-Generation Sequencing, protein analysis and much more.
  
  Diagenode's Bioruptor simultaneously shears multiple samples in sealed tubes of 0.1 ml to 50 ml capacity quickly and accurately, providing optimal fragment lengths. The Bioruptor exhibits a laboratory friendly format as it is easily programmable and capable of processing multiple sample tubes, with parallel processing of 3-12 samples. The instrument is able to sonicate different sample volumes to achieve different fragment size ranges. The closed tube format prevents the sample from cross-contamination and aerosol formation.

Bioruptor 与其他技术的比较

Bioruptor 超声破碎仪与市售的所有其他超声装置相比具有多项优势。