The NEBNext® Microbiome DNA Enrichment Kit facilitates enrichment of microbial DNA from samples containing methylated host DNA (including human), by selective binding and removal of the CpG-methylated host DNA. Importantly, microbial diversity remains intact after enrichment.
Effective enrichment of microbial genomic DNA from samples contaminating host DNA
Fast, simple protocol
Enables microbiome whole genome sequencing, even for samples with high levels of host DNA
Compatible with downstream applications including next generation sequencing on all platforms, qPCR and end point PCR
Suitable for a wide range of sample types
No requirement for live cells
Optional protocol to retain separated host DNA
Also effective for separation of organelle DNA (e.g. mitochondria, chloroplast) from eukaryote nuclear DNA
In human DNA, 4–6% of cytosines are methylated, and 60–90% of these methylated cytosines are at CpG sites (1,2). In contrast, methylation at CpG sites in microbial species is rare. The NEBNext Microbiome DNA Enrichment Kit uses a simple and fast magnetic bead-based method to selectively bind and remove CpG-methylated host DNA. The method uses the MBD2-Fc protein, which is composed of the methylated CpG-specific binding protein MBD2, fused to the Fc fragment of human IgG. The Fc fragment binds readily to Protein A, enabling effective attachment to Protein A-bound magnetic beads. The MBD2 domain of this protein binds specifically and tightly to CpG methylated DNA. Application of a magnetic field then pulls out the CpG-methylated (eukaryotic) DNA, leaving the non-CpG-methylated (microbial) DNA in the supernatant (3). If desired, the host DNA captured in the magnetic bead pellet can be eluted, and a protocol is provided for this.
The NEBNext Microbiome DNA Enrichment Kit is suitable for a wide range of sample types, including samples with high levels of contaminating host DNA, (3,4) and is also effective for separation of organelle DNA (e.g. mitochondria, chloroplast) from eukaryote nuclear DNA (5). The kit is compatible with downstream applications including next generation sequencing on all platforms, qPCR and end-point PCR.
Figure 1. Separation workflow Salivary Microbiome DNA Enrichment DNA was purified from pooled human saliva DNA (Innovative Research) and enriched using the NEBNext Microbiome DNA Enrichment Kit. Libraries were prepared from unenriched and enriched samples and sequenced on the SOLiD 4 platform. The graph shows percentages of 500M-537M SOLiD4 50bp reads that mapped to either the Human reference sequence (hg19) or to a microbe listed in Human Oral Microbiome Database (HOMD)[1]. (Because the HOMD collection is not comprehensive, ~80% of reads in the enriched samples do not map to either database.) Reads were mapped using Bowtie 0.12.7[2] with typical settings (2 mismatches in a 28 bp seed region, etc.).Microbiome Diversity is Retained after Enrichment with the NEBNext Microbiome DNA Enrichment Kit DNA was purified from pooled human saliva DNA (Innovative Research) and enriched using the NEBNext Microbiome DNA Enrichment Kit. Libraries were prepared from unenriched and enriched samples, followed by sequencing on the SOLiD 4 platform. The graph shows a comparison between relative abundance of each bacterial species listed in HOMD[1] before and after enrichment with the NEBNext Microbiome DNA Enrichment Kit. Abundance is inferred from the number of reads mapping to each species as a percentage of all reads mapping to HOMD. High concordance continues even to very low abundance species (inset). We compared 501M 50bp SOliD4 reads in the enriched dataset to 537M 50bp SOLiD4 reads in the unenriched dataset. Reads were mapped using Bowtie 0.12.7[2] with typical settings (2 mismatches in a 28bp seed region, etc).
* Neisseria flavescens – This organism may have unusual methylation density, allowing it to bind the enriching beads at a low level. Other Neisseria species (N. mucosa, N. sicca and N. elongata) are represented, but do not exhibit this anomalous enrichment.
Each kit contains sufficient reagents for the effective separation of CpG methylated DNA from a mixed pool containing microbial or viral DNA. If starting with 1 μg of input DNA per experiment, the volumes provided are sufficient for preparations of up to 6 reactions (NEB #E2612S) and 24 reactions (NEB #E2612L). All reactions should be stored at -20°.
Box 1: Store at -20°C.
Box 2: Store at 4°C. Do not freeze.
This product is related to the following categories:
Use this tool to find citations related to NEBNext products. Search by product, keyword or instrument name – use the filters to select specific product areas.
Publications
George R. Feehery, Erbay Yigit, Samuel O. Oyola, Bradley W. Langhorst, Victor T. Schmidt, Fiona J. Stewart, Eileen T. Dimalanta, Linda A. Amaral-Zettler, Theodore Davis, Michael A. Quail, Sriharsa Pradhan (2013) A Method for Selectively Enriching Microbial DNA from Contaminating Vertebrate Host DNA PLoS One; 8 (10), e76096 . PubMedID: 24204593
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications & Change Notifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
Licenses
MBD-Fc and MCiP was developed by Michael Rehli at the University of Regensburg to improve the sensitivity and specificity of conventional CpG binding techniques.
Limited Use Label License:
This product and its use is the subject of one or more issued and/or pending U.S. and foreign patent applications owned by SEQUENOM, Inc. The purchase of this product from NEB or its authorized distributors conveys to the buyer the non-tranferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for profit entity). The purchase of this product does not convey a license under any method claims in the foregoing patents or patent applications directed to producing the product. The buyer cannot sell or otherwise transfer this product or its components to a third party or otherwise use the product for the following COMMERCIAL PURPOSES: (1) use of the product or its components in manufacturing; (2) use of the product or its components for therapeutic, diagnostic or prophylactic purposes in humans or animals; or (3) resale of the product or its components, whether or not such product or its components are resold for use in research.
ACCEPTANCE OF THE FOREGOING LIMITED USE PROVISIONS: If the purchaser is not willing to accept the limitations of this limited use statement, NEB is willing to accept return of the product with a full refund. For information on purchasing a license to use this product for purposes other than those permitted above, contact the NEB Licensing Department at busdev@neb.com.
The MBD-Fc technology is licensed exclusively to New England Biolabs, Inc. from Sequenom, Inc.
Trademarks
NEW ENGLAND BIOLABS® and NEBNext® are registered trademarks of New England Biolabs, Inc. ULTRA™ is a trademark of New England Biolabs, Inc.
AGENCOURT® and AMPure® are registered trademarks of Beckman Coulter, Inc. ILLUMINA® is a registered trademark of Illumina, Inc. NANODROP® is a registered trademark of Nanodrop Technologies, Inc. QIAGEN® is a registered trademark of The Qiagen Group. ION TORRENT™ and SOLID™ are trademarks of Life Technologies, Inc.
Ineligible item added to cart
Based on your Freezer Program type, you are trying to add a product to your cart that is either not allowed or not allowed with the existing contents of your cart. Please review and update your order accordingly. If you have any questions, please contact Customer Support at freezers@neb.com or 1-800-632-5227 x 8.
Choose your country
North America
Europe
Asia-Pacific
Bienvenue sur notre nouveau site
Chez New England Biolabs, notre objectif a toujours été d’accompagner la communauté scientifique en proposant des produits de haute qualité, une expertise technique et des ressources pédagogiques. Nous sommes ravis d’annoncer le lancement de notre nouveau site web NEB France, conçu pour faciliter l’accès aux contenus scientifiques, aux protocoles, aux ressources techniques et aux informations sur les produits.
Bien que l'expérience utilisateur du site ait été modernisée, notre processus de commande en ligne reste inchangé. Vous pouvez continuer à passer commande via notre boutique en ligne existante en utilisant vos identifiants et numéros de compte habituels.
Nous sommes convaincus que ces améliorations vous permettront de trouver plus facilement les informations dont vous avez besoin et amélioreront votre expérience de navigation.
Si vous rencontrez des difficultés ou souhaitez nous faire part de vos commentaires, n'hésitez pas à nous contacter à l'adresse customersupport.fr@neb.com.
You have been idle for more than 20 minutes, for your security you have been logged out. Please sign back in to continue your session.
Please sign in to continue shopping.
Institution Changed
Your profile has been mapped to an Institution, please sign back for your profile updates to be completed.
Sign in to your NEB account
To save your cart and view previous orders, sign in to your NEB account. Adding products to your cart without being signed in will result in a loss of your cart when you do sign in or leave the site.
Want to try this product? Request a sample
Need a custom/large volume order? Contact us
