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Monarch® HMW DNA Extraction Kit for Cells & Blood

Catalog # Concentration Size List Price Quantity Your Price
T3050L 50 preps $639.00
$575.10
T3050S 5 preps $117.00
$105.30
Catalog # Size List Price Your Price
T3050L 50 preps $639.00
$575.10
T3050S 5 preps $117.00
$105.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
  • Designed for extraction for long-read sequencing applications from cells and blood. See related Monarch Genomic DNA Purification Kit (NEB #T3010) for gDNA extraction for general NGS and sequencing (Illumina® and other).
  • Isolate high molecular weight DNA (100 kb to megabase range) for Oxford Nanopore®, PacBio® sequencing
  • Uses unique, innovative technology for fast, user-friendly workflows, only 30-60 minutes for extraction compared to hours or days.
  • Includes RNase A and Proteinase K for effective RNA removal and digestion of cellular proteins
  • Obtain best-in-class yields of highly-pure and intact DNA
  • For HMW gDNA extraction from tissues, bacteria, and more for long-read sequencing, see related Monarch HMW DNA Extraction Kit for Tissue (NEB #T3060)

Featured Resources:

  • Check out our publication in Biotechniques outlining a simple approach to accurate quantitation of ultra high molecular weight (UHMW) DNA.
  • Review our technical note examining the use and comparison of a novel method to isolate genomic DNA from bacteriophages.
Featured Videos
View Video Library
  • Monarch_MiniprepProtocol_VideoThumb
    Monarch® HMW DNA Extraction from Cell & Blood: Protocol Overview
  • Monarch_MiniprepProtocol_VideoThumb
    Overview of High Molecular Weight DNA Extraction using the Monarch® HMW DNA Extraction Kit for Cells & Blood
  • Monarch_MiniprepProtocol_VideoThumb
    Tips for HMW DNA Extraction from Blood Samples with the Monarch® kit (NEB #T3050)
  • Monarch_MiniprepProtocol_VideoThumb
    Fast and Efficient Extraction of High Molecular Weight DNA 

The Monarch HMW DNA Extraction Kit for Cells & Blood provides a rapid and reliable process for extracting high molecular weight (HMW), intact genomic DNA from cultured cells and whole blood. Utilizing an optimized process that combines gentle cell lysis with a tunable fragment length generation followed by precipitation of the extracted DNA onto the surface of large glass beads, the prep proceeds rapidly and utilizes standard laboratory equipment. DNA size ranges from 50-250 kb for the standard protocol and into the Mb range when the lowest agitation speeds are used. Purified DNA is recovered in high yield with excellent purity, including nearly complete removal of RNA. For cells, the process time is only 30 min, while blood samples require erythrocyte lysis and are processed in 60 min. Purity ratios of 1.80-1.90 and 2.2-2.5 (A260/A280 and A260/A230, respectively), are easily and reproducibly achievable, and purified HMW DNA is suitable for a variety of downstream applications including long-read sequencing (Oxford Nanopore Technologies® and Pacific Biosciences®), optical mapping (Bionano Genomics®), and linked-read genome assembly.

Validated Sample Types

Cells

  • K293
  • HeLa
  • NIH3T3
  • Jurkat
  • K562 (suspension cells)
  • HCT116
  • A549
  • U5Os
  • HepG2
  • NCI-460
  • SK-N-SH
  • Aa23

Blood

  • Human
  • Mouse
  • Rat (fresh only)
  • Rabbit

 

 

  • Pig
  • Horse
  • Cow
  • Rhesus money
  • Goat (fresh only)
  • Sheep (fresh only)
  • Chicken
  • Turkey

Visit our input guidelines or more information

 

 

Figure 1: Workflow for isolation of HMW DNA from cells using the Monarch HMW DNA Extraction Kit






Figure 2: Workflow for isolation of HMW DNA from blood using the Monarch HMW DNA Extraction Kit






Figure 3: Reproducible Extraction of HMW DNA from Cells and Blood with the Monarch HMW DNA Extraction Kit




DNA extracted with Monarch HMW DNA Extraction Kit for Cells & Blood. 1 x 106 fresh HEK293 cells and 500 µl fresh human blood were used as inputs and for preps performed according to the kit instructions using the agitation speed indicated above the gel lanes. 500 ng of DNA from the replicates was resolved by PFGE (1% agarose gel, 6 V/cm, 13°C for 20 hours, switch times ramped from 0.5–94 seconds on a BioRad® CHEF-DR® III System). Yield and purity ratios of the individual preps are shown in the accompanying tables. Lambda PFG Ladder (NEB #N0341) was used as molecular weight standard.





Figure 4: Use of varying agitation speeds during lysis produces tunable fragment length of extracted HMW genomic DNA from cells and blood




Preps were performed on duplicate aliquots of 1 x 106 HEK 293 cells and 500 ìl fresh human blood. Samples were agitated at the indicated speed during the lysis step to control the fragmentation of the DNA. Equal amounts of DNA from the replicates (cells: 500 ng; blood: 650 ng) were resolved by PFGE (1% agarose gel, 6 V/cm, 13°C for 20 hours, switch times ramped from 0.5–94 seconds on a BioRad CHEF-DR III System). Yield and purity ratios of the individual preps are shown in the accompanying tables. Lambda PFG Ladder and Lambda DNA-Hind III Digest (NEB #N0341 and #N3012) were used as molecular weight standards. Yield, purity ratios and DINs of the individual preps are shown in the accompanying tables.





Figure 5: Linear correlation between DNA yield and input for cell and blood samples




Summarized yield data for HMW DNA preps are shown carried out at 2,000 rpm during lysis, using HEK293 cultured cells and fresh human blood samples from different donors as input material in the corresponding protocols. The starting materials were diluted to 5 different concentrations to cover the entire recommended input range. Cell samples ≤ 5x105 cells and blood samples <500 µl were purified using the recommended volumes for low input samples. Obtained yields show a high degree of linearity over the displayed input range.





Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Monarch® RBC Lysis Buffer T3051-21 25 1 x 15 ml
  Monarch® gDNA Nuclei Prep Buffer T3052-21 25 1 x 1.5 ml
  Monarch® gDNA Nuclei Lysis Buffer T3053-21 25 1 x 1.5 ml
  Monarch® Precipitation Enhancer T3055-21 25 1 x 1 ml
  Monarch® gDNA Wash Buffer T3015-21 25 1 x 4.5 ml
  Monarch® gDNA Elution Buffer II T3056-21 25 1 x 1.5 ml
  Proteinase K, Molecular Biology Grade P8200GVIAL -20 1 x 0.11 ml 20 mg/ml
  Monarch® RNase A T3018-21 -20 1 x 0.035 ml 20 mg/ml
  Monarch® DNA Capture Beads T3005-21 25 1 x 10 beads
  Monarch® 2 ml Tubes T3003-21 25 1 x 10 tubes
  Monarch® Bead Retainers T3004-21 25 1 x 5 items
  Monarch® Spin Collection Tubes T2118-23 25 1 x 5 tubes
  Monarch® gDNA Nuclei Prep Buffer T3052-1 25 1 x 10 ml
  Monarch® gDNA Nuclei Lysis Buffer T3053-1 25 1 x 10 ml
  Monarch® Precipitation Enhancer T3055-1 25 1 x 5 ml
  Monarch® gDNA Wash Buffer T3015-1 25 1 x 18 ml
  Monarch® gDNA Elution Buffer II T3056-1 25 1 x 12 ml
  Proteinase K, Molecular Biology Grade P8200AAVIAL -20 1 x 0.55 ml 20 mg/ml
  Monarch® RBC Lysis Buffer T3051-1 25 1 x 160 ml
  Monarch® RNase A T3018-1 -20 2 x 0.17 ml 20 mg/ml
  Monarch® DNA Capture Beads T3005-1 25 1 x 100 beads
  Monarch® 2 ml Tubes T3003-1 25 2 x 50 tubes
  Monarch® Bead Retainers T3004-1 25 1 x 50 items
  Monarch® Spin Collection Tubes T2118-1 25 1 x 50 tubes
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

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Quality Control Assay
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.
Change Notifications

Effective March 18,2024, collection tube dimensions modified and component numbers changed.

Effective March 14, 2024: Proteinase K, Molecular Biology Grade component number has changed with the concentration changing from 800U/ml to 20 mg/ml.

 

Specifications
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]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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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.

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.

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information about commercial rights, please email us at gbd@neb.com While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
 

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