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This product has been discontinued and has been replaced by the MethylCollector MBD Capture Kit, cat# 55026
Active Motif's MethylCollector™ Ultra offers a fast magnetic assay capable of efficiently isolating methylated CpG islands from fragmented genomic DNA*. MethylCollector Ultra is based on the Methylated CpG Island Recovery Assay (MIRA) which uses a MBD2b/MBD3L1 protein complex for improved enrichment of CpG dinucleotides. Enriched DNA can be used in many downstream applications, such as end point or real time PCR analysis of the methylation status of particular loci in normal and diseased samples, bisulfite conversion followed by cloning and sequencing, or amplification and labeling for microarray analysis.
To learn more about how the kit works, please select the Description tab below. To see data and results, including comparisons to alternative methods, select the Data tab below.
Publication: A Versatile Assay for Detection of Aberrant DNA Methylation in Bladder Cancer. S Tommasi, A Besaratinia - Urothelial Carcinoma, 2018 - Springer
*Technology covered under U.S. Patent No. 7,425,415.
| Name | Format | Cat No. | Price | |
|---|---|---|---|---|
| MethylCollector™ Ultra | 30 rxns | 55005 | Discontinued | |
| MethylCollector™ Ultra Manual |
| MethylCollector & MethylCollector Ultra-Seq MOTIFvations June 13 Article |
| Products and Services to Study DNA Methylation |
| Epigenetics Products and Services |
Preparing DNA Samples for Next-Gen Sequencing following MethylCollector Ultra Enrichment
Enriched methylated DNA obtained using MethylCollector Ultra can be used directly in Next-Gen Sequencing sample processing protocols.
If you intend to perform Next-Gen sequencing analysis following MethylCollector Ultra enrichment of methylated DNA, please refer to the MethylCollector Ultra manual and the Illumina ChIP-Seq DNA Sample Prep Kit for instructions on how to prepare your DNA samples for Next-Gen sequencing.
Click on image to enlarge size.
Figure 1: Next-Gen sequencing data generated using MethylCollector Ultra enriched DNA correlates well with CpG density.
DNA was enriched from 1 µg of human PBMC DNA using Active Motif’s MethylCollector Ultra Kit. Next-Gen sequencing was performed using the Illumina platform to generate 28 million sequence tags. Tags were mapped to generate a whole-genome DNA methylation profile. The image above shows that the enriched regions (purple peaks) correlate well with CpG density except at CpG islands, which should be largely unmethylated.
Click on image to enlarge size.
Figure 2: Next-Gen sequencing data generated using MethylCollector Ultra enriched DNA is more sensitive and robust than data from other MBD enrichment kits.
DNA was enriched from 1 µg of human PBMC DNA using Active Motif’s MethylCollector Ultra Kit and a competitor’s MBD-based enrichment kit. Next-Gen sequencing was performed using the Illumina platform to generate 28 million sequence tags. Tags were mapped to generate whole-genome DNA methylation profiles. The top purple panel is data produced from MethylCollector Ultra and the orange panel is data from the Competitor M kit. MethylCollector Ultra identified more methylated regions and the peaks had greater intensity than peaks identified from the competitor kit.
Click on image to enlarge size.
Figure 3: Next-Gen sequencing data generated using MethylCollector Ultra detects methylation at CpG shores.
DNA was enriched from 1 µg of human PBMC DNA using Active Motif’s MethylCollector Ultra Kit. Next-Gen sequencing was performed using the Illumina platform to generate 28 million sequence tags. Tags were mapped to generate a whole-genome DNA methylation profile. The image above shows one of many examples of DNA methylation being detected at CpG shores rather than in the CpG island itself. This data agrees with recent findings showing that a large portion of methylated sites occur in these CpG shore regions that are adjacent to CpG islands.
