Chromatin Preparation From Tissues for Chromatin Immunoprecipitation Using Diagenode’s Chromatin Shearing Optimization Kit - Low SDS and Bioruptor®

29 Oct 2014

This application note describes the chromatin preparation from fresh or frozen tissues. The isolated chromatin can be used for chromatin immunoprecipitation assays using Diagenode’s iDeal ChIP-seq kit (C01010051). The following protocol is optimized for 30-40 mg of tissue allowing up to 18 ChIP samples (1.5-2 mg of tissue per sample). However, the exact amount of tissue needed for ChIP may vary depending on protein abundance, antibody affinity etc. and should be determined for each tissue type.

Bioruptor® Pico

Diagenode

The Bioruptor® Pico is the new next-generation all-in-one shearing device, highly optimized for DNA shearing for next-generation sequencing, chromatin shearing for ChIP-seq applications, RNA shearing, protein extraction, and cell/tissue lysis. The Pico is the shearing device of choice for sequencing applications, providing optimal yields, lengths, and consistency and excels in shearing different fragment size ranges, which are frequently required for downstream applications (e.g. bridge amplification) for sequencing. Moreover, the Pico provides flexibility in other applications saving the need to buy additional equipment. Unlike competing systems, the Pico has high throughput capabilities and can process up to 12 samples at once. The only ultrasonicator you’ll ever need, optimized for: Next-Generation Sequencing (5 - 100 µl) Chromatin shearing (10 µl - 2 ml) RNA shearing Protein extraction Tissue and cell lysis Simultaneous sonication of 12 samples Small and light  

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Bioruptor® Plus

Diagenode

The Bioruptor® uses a unique system to uniformely process multiple samples in sealed tubes of 0.5 ml to 50 ml capacity. An excellent device for shearing chromatin or DNA, cell and tissue disruption and many other applications. 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. Features: Digital timer Metal soundproof box Overheat shutdown protection (this feature is not available in the US) Machine lifetime control Applications: Chromatin Shearing DNA Shearing Cell & Tissue disruption

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iDeal ChIP-seq kit

Diagenode

The iDeal ChIP Kit for Transcription Factors.Diagenode’s iDeal ChIP-seq Kit for Transcriptions Factors is a highly specialized solution for robust ChIP-seq results transcription factors. Unlike competing solutions, our kit utilizes a highly optimized protocol and is backed by validation with a broad number and range of transcription factors. The kit provides high yields with excellent specificity and sensitivity. Advantages over competing solutions: Confidence in results: Validated for ChIP-seq with multiple transcription factors Proven: Validated by the epigenetics community, including the BLUEPRINT consortium Most complete kit available for highest quality data -- includes controls, primers, and magnetic beads Validated with Diagenode’s MicroPlex Library Preparation Kit and IP-Star Automation Completely automatable

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Chromatin Preparation From Tissues for Chromatin Immunoprecipitation Using Diagenode’s Chromatin Shearing Optimization Kit - Low SDS and Bioruptor®