June 2013 • volume 14 • number 2

DNA Methylation

MethylCollector™ Ultra for the Most Sensitive Enrichment of Methylated DNA
Active Motif’s popular MethylCollector™ Ultra Kit is the fastest, most sensitive and speciﬁc kit available for enrichment of CpG methylated dinucleotides from fragmented genomic DNA. The patented MIRA (Methylated CpG Island
Recovery Assay) technology combined with a magnetic bead-based protocol enables the most efﬁcient methylated
DNA enrichment method possible, with results in less than half the time required for MeDIP.
MethylCollector Ultra for the fastestenrichment of methylated DNA
Active Motif’s MethylCollector Ultra Kit
provides the most rapid and sensitive
method for enrichment of methylatedDNA. The kit’s design is based on thepatented MIRA technology that uses apuriﬁed MBD2b/MBD3L1 protein complex to bind methylated DNA. This highafﬁnity binding method provides greater
enrichment than other MBD capture orantibody immunoprecipitation (MeDIP)
methods (Table 1). MethylCollector Ultrarequires as little as 1 ng (~200 cells) ofinput DNA and enables detection ofmethylation from DNA fragments
containing as few as 5 methylated CpGs.
Magnetic capture allows more rapid andefﬁcient recovery of methylated DNA.
Recovered dsDNA is compatible withvarious downstream applications including Next-Gen Sequencing (Figure 1).

MethylCollector™ Ultra (Cat. No. 55005)

MeDIP Kit (Cat. No. 55009)

Input material: 1 ng - 1 µg

Input material: 100 ng - 1 µg

–Denature DNA for 10 minutes at 95°C

Combine DNA with MBD2b/MBD3L1 protein mix
and nickel-coated magnetic beads for 1 hour at 4°C

Combine DNA, antibody, bridging antibody andmagnetic beads, incubate overnight at 4°C

Wash beads 4 times using the included magnet

Wash beads 5 times

Elute methylated DNA at 50°C for 30 minutes

Elute methylated DNA at 4°C for 15 minutes

Add Proteinase K Stop Solution

Add Neutralization Buffer

DNA clean up by phenol/chloroform and ethanol
precipitation, or with puriﬁcation columns

DNA clean up by phenol/chloroform and ethanol
precipitation, or with puriﬁcation columns

DNA is ready for analysis

DNA is ready for analysis

Total time required: < 3 hours

Total time required: > 7 hours

Table 1: Comparison of the MethylCollector™ Ultra process to MeDIP.

NEW: MethylCollector Ultra-Seq Services Now Available
Active Motif has now expanded its
offerings to include end-to-end MethylCollector™ Ultra-Seq Epigenetic Services.

Simply submit your cells or DNA andreceive fully analyzed genome-wide
methylation data in just 6-8 weeks.

MIRA-Seq Service includes:• DNA isolation from cells or tissues• Methylated DNA enrichment using
MethylCollector Ultra• qPCR analysis of positive andnegative control sites• Next-Gen Library generation• Sequencing of at least 30 million
tags using the Illumina HiSeqplatform• Analysis: mapping, peak calling,visualization ﬁles and Excel output

MethylCollector Ultra

www.activemotif.com

For complete details on this and otherservices offered, please visit us atwww.activemotif.com/services.

Competitor M

THRAP3

Clorf113
p34.3

36,760,000

36,780,000

36,800,000

36,820,000

STK40

36,840,000

LSM10Figure 1: MethylCollector Ultra is more sensitive than other MBD-based enrichment methods.
The performance of MethylCollector Ultra was compared to that of an alternative MBD capture kit. Methylated DNA wasisolated from 1 µg of human PBMC DNA using both enrichment kits. The enriched DNA was sequenced using the Illumina
HiSeq platform. Regions of enrichment from a 1.2 Mb region of the genome are shown. The MethylCollector Ultra technology
detected more methylated regions and the peaks were of greater intensity than those detected by the competitor’s kit.

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