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Science  11 Oct 2013:
Vol. 342, Issue 6155, pp. 265
DOI: 10.1126/science.342.6155.265-a

DNA ENRICHMENT KITS

PointMan DNA Enrichment kits comprise three kits for enriching mutations in BRAF, KRAS, and EGFR T790M genes associated with skin melanoma, colorectal, and lung cancers. PointMan kits offer highly specific and ultrasensitive enrichment of mutant genes in a background of wild-type (normal) genes that is unmatched by existing technologies. PointMan is a real-time polymerase chain reaction (PCR) technology that provides reliable and extremely sensitive detection for cancer mutations. It is highly efficient in amplifying the target sequence of interest, while suppressing amplification of the wild type. The resulting sample is effectively enriched for the mutation, thereby having the potential to offer industry leading sensitivity in a wide variety of sample types. PointMan DNA enrichment kits can also be used to enrich all mutant sequences within the gene of interest using a single set of reagents, unlike competing technology that requires a separate reagent set for each mutation within a gene sequence.

DNA METHYLATION ANALYSIS

TrueMethyl brings unprecedented clarity to the analysis of DNA by providing quantitative, accurate, and repeatable single-base resolution sequencing of the modified bases hydroxymethylcytosine (5-hmC) and methylcytosine (5-mC) for the first time. Traditional bisulfite sequencing cannot discriminate between 5-hmC and 5-mC. Recent studies have shown that at some sites in the genome the level of 5-hmC can be comparable to the level of 5-mC, emphasizing the importance of identifying these variants accurately. The high-quality and easy-to-use TrueMethyl kits utilize innovative oxidative bisulfite sequencing (oxBS-Seq). TrueMethyl kits can be used with a variety of common platforms including next generation sequencing systems, methylation arrays, and targeted assays. Trials of the kits conducted at leading research centers around the world have already begun to yield new insights into genome function and highlighted the advantages of TrueMethyl relative to traditional approaches.

DIGITAL PCR ASSAYS

PrimePCR assays, for Droplet Digital polymerase chain reaction (ddPCR), are predesigned assays for mutation detection and copy number variation (CNV), which have been experimentally validated to provide single-copy PCR resolution without a standard curve. Droplet Digital PCR technology provides an absolute measure of target DNA molecules. The assays, combined with the sensitivity of Bio-Rad's ddPCR system, are capable of detecting a single mutant copy in a background of 2,000 or more wild-type molecules (0.05% mutation frequency). The precision of ddPCR also enables cancer researchers to discriminate small-fold copy number changes with the 62 assays targeting common cancer genes and two reference target assays now available. The PrimePCR ddPCR assays employ universal cycling conditions and do not require optimization. Primer specificity has been validated by next generation sequencing, and all assays have been validated on Bio-Rad's QX100 Droplet Digital PCR system. PrimePCR ddPCR assays are available in multiple reaction sizes.

GENOME EDITING TOOLS

Sigma CRISPRs is an inexpensive mammalian genome editing tool suitable for screening and exploratory studies. Sigma CRISPRs are packaged in a single plasmid vector containing GFP for fast enrichment of gene-edited cells and can be custom-designed online using an exclusive bioinformatics tool. Derived from bacterial and archaeal immune defenses, which guard against invading viruses and foreign DNA, Sigma CRISPRs utilize customizable RNA—nuclease complexes that allow introduction of genome edits at a desired site in mammalian genomes with high efficiency. Sigma's exclusive online design tool allows users to identify the best target site(s) within human, mouse, and rat protein-coding exons that adhere to CRISPR/Cas targeting requirements. It is the only commercial tool that minimizes the possibility of off-target effects by automatically applying CRISPR/Cas best design practices, including a minimum requirement of three base-pair mismatches between the target site and other sites in the genome.

  • Sigma-Aldrich

  • For info: 800-325-3010

  • www.sigma.com/crisprs

DNA PURIFICATION

MagSi-DNA cleanFIX offers a cost-effective solution for purifying DNA fragments, removing all unwanted side products and reagents. MagSi-DNA cleanFIX enables a combination of polymerase chain reaction (PCR) cleanup and dye terminator removal from sequence reaction mixes with a single product. The kit uses magnetic silica bead technology and is easily automated because no columns or centrifugation steps are involved. The kit uses a simple "Bind, Wash, and Elute" procedure common to magnetic particle purification protocols. The precision and reproducibility of the workstation in combination with the customized magnets for MagSi-DNA cleanFIX allow fast magnetic separation and homogenization of the samples. Overall, the PCR cleanup protocol using MagSi-DNA cleanFIX is shown to enable fast and efficient recovery of DNA fragments larger than 80 bp with >99% removal of primers and primer dimers. In addition, the dye terminator removal method consistently delivered high-quality sequence data with Phred >20 scores above 700.

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  • Newly offered instrumentation, apparatus, and laboratory materials of interest to researchers in all disciplines in academic, industrial, and governmental organizations are featured in this space. Emphasis is given to purpose, chief characteristics, and availabilty of products and materials. Endorsement by Science or AAAS of any products or materials mentioned is not implied. Additional information may be obtained from the manufacturer or supplier.

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