Efficient and reliable shearing of chromatin for high-throughput analysis

-本申请是为了满足染色质免疫沉淀（ChIP）和染色质RNA免疫沉淀（ChRIP）等方案对可靠和可重复剪切DNA的直接和迫切需求。这些技术分别鉴定蛋白质- dna和rna -蛋白质- dna相互作用，并且需要超声（染色质剪切）步骤，这对实验的成功至关重要。所要求的设备是科瓦利斯ME220聚焦超声仪，这将有助于在染色质水平上研究这些相互作用的许多方面。本申请的四名申请人拥有互补的专业知识，并使用广泛的生物系统来解决这些关键问题。他们专注于回答以下生物学问题：1)转录因子和非编码rna的细胞状态稳定（Hussein团队），2)DNA损伤/修复和染色质之间的动力学（fraet - turcotte团队），3)染色体结构和转录调控（Bilodeau团队），4)核糖体rna的生物发生和表观遗传调控（Moss团队）。***科瓦利斯ME220聚焦超声仪提供了几个特性，将提高我们结果的可靠性和质量，并满足我们的大部分需求。它克服了由于过时的超声波产生的不可靠和不一致的结果以及样品和体积限制而引起的许多技术和实际问题。ME220聚焦超声仪由于其高性能的样品制备和自适应聚焦声学设计，提供了可重复的结果。这一特点防止热引起的样品损坏，这可能导致实验之间的偏差和变化。它允许使用8个样品同时与一个附加选项升级到96个样品。这个选项增加了声纳器的多功能性和高吞吐量能力，这在其他声纳器中是没有的。它可以使用低至15µL的体积，这是一个非常重要的功能，当使用珍贵的样品或极低细胞数的样品时。此外，ME220聚焦超声仪是一个用户友好和精心设计的系统，预装协议，使HQP培训从所有水平的经验。总体而言，ME220聚焦超声仪优于市场上许多可用的超声仪，并将取代我们过时的系统。此外，它将提高我们的竞争力和我们为学术和工业就业市场生产高技能HQP的能力。*****

- This application is to finance an immediate and urgent need for reliably and reproducibly shearing DNA for protocols such as Chromatin immunoprecipitation (ChIP) and Chromatin RNA immunoprecipitation (ChRIP). These techniques identify Protein-DNA and RNA-Protein-DNA interactions, respectively, and require a sonication (chromatin shearing) step that is critical for the success of the experiments. The requested equipment is a Covaris ME220 Focused-ultrasonicator that will facilitate the study of many aspects of these interactions at the chromatin level. The four applicants on this application possess complementary expertise and use a broad range of biological systems to address these critical questions. They focus on answering biological questions regarding: 1) cell state stabilization by transcription factors and non-coding RNAs (team Hussein), 2) the dynamics between DNA damage/repair and chromatin (team Fradet-Turcotte), 3) chromosome architecture and transcriptional regulation (team Bilodeau), and 4) the biogenesis and epigenetic regulation of ribosomal RNAs (team Moss).***The Covaris ME220 focused-ultrasonicator provides several characteristics that will improve the reliability and quality of our results and meet much of our needs. It overcomes many of the technical and practical issues that are raised due to unreliable and inconsistent results produced by outdated sonicators, and sample and volume limitations. The ME220 focused-ultrasonicator provides reproducible results due to its high-performance sample preparation and Adaptive Focused Acoustics design. This feature prevents heat induced sample damage, which can cause bias and variation between experiments. It allows using 8 samples simultaneously with an add-on option to upgrade to 96 samples. This option increases the versatility and high-throughput capacity of the sonicator, which is not found in other sonicators. It can use volumes as low as 15µL, which is an extremely important feature, when using precious samples or samples with very low cell numbers. Furthermore, the ME220 focused-ultrasonicator is a user-friendly and well-designed system with preloaded protocols that enable training of HQP from all levels of experience. Overall, the ME220 focused-ultrasonicator outperforms many available sonicators on the market and will replace our outdated system. Moreover, it will increase our competitiveness and our ability to produce highly skilled HQP for the academic as well as industrial job market.*****