Successful Application of Novel Plant ChIP-seq Technology Results in High Quality Sequencing Data from Limited Amounts of <i>Arabidopsis thaliana</i> Seedling Cells

13 Mar 2014

This study presents a successful and reliable ChIP-seq technology using limited amounts of plant material, with specific reagents and protocols, supported by strict bioinformatic QC analysis.

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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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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MicroPlex Library Preparation™ Kit

Diagenode

The easiest NGS Library preparation you will ever find! The MicroPlex Library Preparation™ kit is the only kit on the market which is validated for ChIP-seq and which allows the preparation of indexed libraries from just picogram inputs. In combination with the True MicrochIP kit, it allows for performing ChIP-seq on as few as 10,000 cells. Studies showed a perfect match between the top 40% of peaks and the reference ChIP-seq dataset. Based on the NIH Encode project criterion, ChIP-seq results are considered reproducible between an original and reproduced dataset if the top 40% of peaks have at least an 80% overlap ratio with the compared dataset. With less input, fewer steps, fewer supplies, and faster time to results, the MicroPlex kit is the library preparation solution of choice. MicroPlex Library Preparation™ Kit Features: Simplified protocol: single tube, 3-step protocol without any intermediate purifications 3 times faster than TruSeq™ - only 2 hours for entire protocol DNA sample inputs from only 50 pg Specifically optimized for ChIP-seq Fewer PCR cycles: less bias Indexing options, easily automated Proven accuracy and reproducibility  Diagenode’s MicroPlex Library Preparation Kits v3 have been extensively validated for ChIP-seq samples and are optimized to generate DNA libraries with high molecular complexity from the lowest input amounts – down to 50 pg. The kit MicroPlex v3 includes all buffers and enzymes necessary for the library preparation. For flexibility of the choice different formats of compatible dual indexes are available separately.  

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Genome AnalysisGenomics, the study of genomes, includes functional genomics, evolutionary genomics and comparative genomics. There are many genomic technologies such as DNA sequencing of whole genomes, computational biology and bioinformatics. DNA and nucleic acids must be isolated and concentrated from cells for analysis with kits, automated analyzers and software. Other useful technologies for studying genomics include PCR, microarrays and electrophoresis.DNA / RNA Extraction and PurificationPurified DNA and RNA are required for numerous downstream molecular biology applications. Consequently, the importance of high-quality DNA/RNA extraction and purification equipment cannot be underestimated. Many purification kits are available and are typically optimized for nucleic acid type and source, including plasmid DNA, genomic DNA, mRNA, RNA and viral nucleic acid purification kits. Automated extraction and purification of nucleic acids can be implemented with magnetic bead separator instruments or high-throughput purification workstations. Find the best DNA/RNA extraction and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.DNA SequencingDNA sequencing, such as sanger sequencing, is a biological technique that determines the precise order of nucleotide bases in a fragment or template of DNA. DNA sequencers and genetic analyzers are based on capillary electrophoresis, where labeled DNA fragments are electrophoretically separated by size as they migrate through a polymer. Find the best DNA sequencing products, including DNA sequencing kits, genomic libraries and genetic identity kits in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Next Generation SequencingNext-generation sequencing (NGS), also known as whole-genome sequencing, high-throughput sequencing and massive parallel sequencing, produces and analyses thousands to millions of nucleotide sequences at once. Sequencing systems operate via varying technologies depending on the manufacturer, including sequencing by synthesis, ligation, pyrosequencing, ion semiconductor and single-molecule real-time sequencing. For NGS, library preparation is paramount to successful sequencing. In this section, explore a range of library preparation kits, from targeted, amplicon-based or hybridization-based kits including epigenomic, transcriptomic and genomic workflows to fragmentation kits. Find the best next-generation sequencing products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Epigenetic AnalysisEpigenetic analysis products are used to study heritable phenotype changes that do not alter the DNA sequence, such as DNA modifications affecting gene activity. Determine whether genes have been methylated with DNA methylation quantification kits or methylation microarrays. Assess epigenetic regulation with histone modification kits or ChIP-on-chip microarrays. Kits are also available for bisulfite conversion and amplification of your samples, as well as epigenomic library preparation kits for next-generation sequencing (NGS). Find the best epigenetic products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.ChIP-SequencingBioinformaticsBioinformatics uses computational models and methodologies to study biological processes and to understand biological data.Plant SciencePlant science studies the biology of plants, including genetics, physiology, ecology, and biotechnology. Research in this field is essential for improving crop yields, disease resistance, and sustainability. Explore plant science tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.