The Monarch Spin High-Capacity DNA Cleanup Kit (100 μg) offers class-leading binding capacity and proven Monarch buffer chemistry, while maintaining the convenient spin column format for silica membrane-based DNA cleanup. This is a high-capacity version of our Monarch Spin PCR & DNA Cleanup Kit (5 μg) (NEB #T1130). The Monarch Spin High-Capacity DNA Cleanup Kit (100 μg) efficiently purifies and concentrates up to 100 μg of high-quality, double-stranded and single-stranded DNA from enzymatic reactions such as RCA, RE digestion, labeling, or exonuclease treatment. The high capacity and rapid process time make the kit an excellent choice for cleanup of templates generated by cell-free DNA synthesis workflows. Modified protocols enabling cleanup of 100 µg amounts of oligonucleotides, genomic DNA, or RCA products make this kit a workhorse for multiplexed, high-throughput workflows requiring large amounts of starting materials. The kit features precision-engineered spin columns uniquely designed to enable low elution volumes (50 μl), resulting in highly concentrated DNA suitable for downstream applications such as in vitro transcription (IVT), genome editing, or viral vector transfection.
The kit includes spin columns, collection tubes, buffers, and other reagents, all compactly packaged. The spin columns are compatible with microcentrifuges, centrifuges, and vacuum manifolds, without additional plastic accessories. Manufactured using state-of-the-art automation, Monarch spin columns ensure high quality and consistency. The optimized buffer system helps ensure efficient DNA binding and purification, without the need to adjust pH.
Monarch kits are more eco-friendly than other options from leading suppliers, reducing plastic usage in the columns and bottles while maintaining structural integrity for centrifugation or vacuum manifolds.
Properties
Purification format
Spin column
DNA Sample
DNA from PCR and other enzymatic reactions (e.g., restriction digests, RCA, labeling, exonuclease treatment, ligations)
ssDNA or dsDNA oligonucleotides from enzymatic reactions can also be purified using the Oligonucleotide Cleanup Protocol
Genomic DNA and large size DNA (>25 kb) or RCA reaction cleanup using the supplemental protocols
Typical Recovery
70-90%
DNA Purity
A260/280 > 1.8 and A260/230 > 1.8
Nucleic Acid Binding Capacity
Up to 100 μg of DNA
Elution Volume
50-200 μl
DNA Size Range
Standard protocol: 40 bp – 25 kb
Oligonucleotide Cleanup protocol: ssDNA >12 nt and dsDNA >10 nt
Protocol Time
9 minutes of spin and incubation time
Compatible Downstream Applications
Ligation, in vitro transcription (IVT), genome editing, viral transfection, restriction digestion, labeling and other enzymatic manipulations, library construction and DNA sequencing
Figure 1: Monarch Spin High-Capacity DNA Cleanup Kit (100 µg) offers a wide range of DNA input amounts
DNA purifications were performed using the Monarch Spin High-Capacity DNA Cleanup Kit (100 µg) with varying amounts of DNA. To assess average DNA recovery, DNA input (1 kb Ladder, NEB #N3232) ranging from 10 to 100 µg was used and eluted in 100 µl of Monarch Buffer EY, in triplicate. DNA concentrations for both input and eluted samples were measured using a Trinean DropSense 16. Percent recovery calculations were based on the measured DNA concentrations and the elution volume.
Figure 2: Monarch Spin High-Capacity DNA Cleanup Kit (100 µg) effectively recovers various DNA sample types, maximizing utility of the kit
DNA samples were purified in triplicate using the Monarch Spin High-Capacity DNA Cleanup Kit (100 µg), following either the standard protocol (for linear, circular, and ssDNA) or the supplemental protocol (for large DNA). Each sample, except for the RCA product (50 µl, 1 mg/ml), contained 10 µg of DNA and was eluted in 100 µl of Monarch Buffer EY. DNA concentrations for both input and eluted samples were measured using a Trinean DropSense 16, except for the RCA product, which was measured using the Qubit dsDNA Broad Range (BR) Assay Kit. Percent recovery was calculated based on the measured DNA concentrations and elution volumes, with results displayed in a bar graph. For linear DNA, the following ladders were utilized: LMW DNA ladder (25-766 bp, NEB #N3233), 1 kb DNA Ladder (0.5–10 kb, NEB #N3232), and HindIII digest (2–23 kb, NEB #N3012). For circular DNA, pUC19 (NEB #N3041), PhiX174 RF I (NEB #N3021) and PhiX174 RF II (NEB #N3022) were employed. For ssDNA, phiX174 Virion (NEB #N3023) and synthetic ssDNA (150 nt) were used.
Figure 3: Monarch Spin High-Capacity DNA Cleanup Kit (100 µg) effectively recovers oligonucleotide DNA using the oligonucleotide cleanup protocol
DNA samples were purified in triplicate using the Monarch Spin High-Capacity DNA Cleanup Kit (100 µg) with the provided oligonucleotide cleanup protocol. To assess average oligonucleotide recovery, 10 µg of synthesized ssDNA (8–25 nt) and dsDNA (8–25 bp) were used and eluted in 100 µl of Monarch Buffer EY. DNA concentrations for both input and eluted samples were measured using a Trinean DropSense 16. Percent recovery was calculated based on the measured DNA concentrations and elution volumes, with results displayed in a bar graph. Using this protocol, ssDNA (≥ 12 nt) and dsDNA (10 bp) can be effectively recovered.
Figure 4: The Monarch Spin High-Capacity DNA Cleanup Kit (100 µg) efficiently removes Proteinase K, demonstrating protein removal from enzymatic reactions
DNA samples containing 0.8 units or 8 units of Proteinase K (NEB #P8200SVIAL) were purified using the Monarch Spin High-Capacity DNA Cleanup Kit (100 µg). The eluted DNA was then tested for residual Proteinase K by incubating 10 µg MBP5 in 1X NEBuffer 4 at 37°C for 5 minutes. A proteinase K control titration (0.002-3.12 x10-5 units, 2-fold dilution, and 0 units) was used as a control, as shown in the left gel. The right gel represents six replicate purified samples with 0.8 units or 8 units of Proteinase K in the input sample, showing that MBP5 remains undigested at 42.5 kDa.
Figure 5: DNA purified with the Monarch Spin High-Capacity DNA Cleanup Kit (100 µg) is of high quality and ideal for downstream applications
To demonstrate downstream application compatibility, an in vitro transcription (IVT) reaction was performed using a DNA template purified with the Monarch Spin High-Capacity DNA Cleanup Kit (100 µg). The pCMV-CLuc 2 plasmid or RCA product, using pCMV-CLuc 2 as a template, was linearized with NotI-HF (NEB #R3189). The linearized DNA was then purified using the Monarch Spin High-Capacity DNA Cleanup Kit (100 µg), resulting in highly pure DNA, as indicated by the A260/230 and A260/280 ratios. This purified DNA served as the template for the IVT reaction, using the HiScribe® T7 High Yield RNA Synthesis Kit (NEB #E2040). The reaction was scaled to 100 µl, with a FLuc IVT reaction performed as a control using the FLuc Control Template provided in the HiScribe T7 High Yield RNA Synthesis Kit (NEB #E2040).
Figure 6: Overview of the Monarch Spin High-Capacity DNA Cleanup Kit (100 µg) workflow
Figure 7: Features of the Monarch Spin Column S3A for High-Capacity DNA Cleanup
This product is related to the following categories:
High Performance: Achieve high yields (up to 100 μg) and high purity in the purification, cleanup, and concentration of DNA, with the capability to remove short primers, detergents, and other low-molecular-weight reaction components (e.g. nucleotides, DMSO, betaine).
High Concentration: Elute in very small volumes, in as little as 50 μl for elution, allowing for highly concentrated DNA.
High Compatibility: Designed for maximum compatibility with microcentrifuges and vacuum manifolds using a spin column format. No extra accessories or plastics required.
Broad Utility: Purify large (> 25 kb) or small DNA fragments (< 40 bp), including oligonucleotides, with this single kit, using the modified protocols provided.
Unique Design: Spin column features a unique design that enables elution in low volumes and minimizes buffer retention and contaminant carryover.
Optimized: Buffer system is optimized without the need to adjust pH.
Application Compatibility: Purified DNA is ready for downstream molecular applications, such as in vitro transcription template, restriction digests, DNA sequencing, ligation, amplification, and other enzymatic reactions.
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]
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]
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.
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