The Monarch Mag PCR & DNA Cleanup Kit (5 μg) enables rapid and reliable purification and concentration of up to 5 μg of DNA, including both single- and double- stranded forms, from common enzymatic reactions such as PCR, restriction digestion, ligation, reverse transcription, and rolling circle amplification (RCA). Using a streamlined bind-wash-elute workflow with silica-coated magnetic beads, the kit delivers highly pure, inhibitor-free DNA ready for immediate use in downstream applications such as in vitro transcription (IVT), sequencing, restriction digestion, library construction, labeling, and other enzymatic manipulations. The magnetic bead format offers flexibility and scalability, supporting both manual and automated workflows. Additional modified protocols support varying DNA input amounts and sample volumes and enable purification of oligos, genomic DNA, and RCA products.
The kit includes magnetic beads, optimized buffers, and reagents, all compactly packaged. The magnetic bead technology in this kit eliminates the need for centrifugation or vacuum-based processing and requires no pH monitoring, with the optimized buffer chemistry.
Monarch kits are manufactured to ensure high quality and consistency. As with all Monarch kits, this product is designed with sustainability in mind, reducing plastic usage where possible and in the design of buffer bottles and other plastic components, while maintaining performance and reliability across a wide range of applications.
Properties
Purification Format
Magnetic bead
DNA Sample Type Compatibility
DNA from various enzymatic reactions (restriction digestion, ligation, reverse transcription, PCR, RCA, etc.)
dsDNA, ssDNA, circular/linear DNA, oligonucleotides, gDNA, and RCA
Typical Recovery
70 - 90%
DNA Purity
A260/280 > 1.8 and A260/230 > 1.8
Binding Capacity
Up to 5 µg using a standard protocol. The binding capacity can be scaled up or down as needed to accommodate different sample requirements.
Elution Volume
25–50 µl
DNA Size Range
Standard protocol: 50 bp – 25 kb
Oligonucleotide Cleanup protocol: ssDNA ≥18 nt and dsDNA ≥14 nt
gDNA cleanup protocol: ≥25 kb
Compatible Downstream Applications
Ligation, restriction digestion, labeling, and other enzymatic manipulations, cloning library construction, in vitro transcription template preparation, and general DNA sequencing.
Figure 1: Automated PCR cleanup with Monarch Mag PCR & DNA Cleanup Kit (5 µg) yields efficient and consistent results
Automated PCR cleanup with Monarch Mag PCR & DNA Cleanup Kit (5 µg) (NEB #T4130) yields efficient and consistent results. PCR cleanup was demonstrated by performing 96 identical 50 μl reactions using Q5® Hot Start High-Fidelity 2X Master Mix (NEB #M0494) with primers amplifying a 673-bp fragment from genomic DNA. PCR products were purified with the Monarch Mag PCR & DNA Cleanup Kit (5 μg) on the KingFisher® Flex instrument. Purified DNA quality was assessed by Trinean DropSense 16 and NanoDrop® 8000, with A260/280 and A260/230 ratios > 1.8 indicating minimal salt and protein presence in the eluted DNA. Integrity of the amplicon was confirmed by resolution on a 1.2% agarose gel, showing a single band of the expected size (L: 100 bp DNA Ladder, NEB #N3231).
Figure 2: Monarch Mag PCR & DNA Cleanup Kit (5 µg) enables versatile purification of diverse DNA types
Monarch Mag PCR & DNA Cleanup Kit (5 µg) (NEB #T4130) enables versatile purification of diverse DNA types. DNA samples were purified in triplicate using the Monarch Mag PCR & DNA Cleanup Kit (5 µg) with a manual workflow, following either the standard protocol (for linear, circular and single-stranded DNA) or the supplemental protocol (for large DNA fragments, oligonucleotides and RCA products). Each sample contained 1 µg of DNA and was eluted in 30 µl of Monarch Buffer EY, except for the RCA product (20 µl at 1 mg/ml). DNA concentrations for both input and eluted samples were measured using the Trinean DropSense 16, except for the RCA product, which was quantified using the Qubit® dsDNA Broad Range Assay Kit. Percent recovery was calculated based on measured DNA concentrations and elution volumes, and the results are presented in a bar graph.
DNA sources included: LMW DNA Ladder (50–1,350 bp, NEB #N3236), 1 kb DNA Ladder (0.5–10 kb, NEB #N3232), HindIII Digest (2–23 kb, NEB #N3012), Supercoiled DNA Ladder (2–10 kb, NEB #N0472), PhiX174 RF I (NEB #N3021), M13mp18 Single-Stranded DNA (NEB #N4040S), PhiX174 Virion (NEB #N3023), Lambda DNA (NEB #N3011), Genomic DNA extracted from HEK293T cells, Synthetic oligonucleotides (16-mer and 20-mer), RCA reaction product.
Figure 3: Monarch Mag PCR & DNA Cleanup Kit (5 µg) provides superior DNA recovery across a range of DNA sizes compared to a leading supplier
Monarch Mag PCR & DNA Cleanup Kit (5 µg) (NEB #T4130) provides superior DNA recovery across a range of fragment sizes compared to a leading supplier. This kit is designed for DNA cleanup without size selection capabilities, retaining all fragment sizes within 50bp - 25 kb. DNA purifications were performed and compared using the Monarch Mag PCR & DNA Cleanup Kit (5 µg) and the leading supplier’s product. To assess recovery across various fragment sizes, 1 µg of a LMW DNA ladder (25–766 bp, NEB #N3233), 1 kb DNA Ladder (0.5–10 kb, NEB #N3232), Lambda DNA HindIII Digest (2–23 kb, NEB #N3012), and 5 µg of a 3 kb fragment were used. DNA concentrations of both input and eluted samples were measured using a Trinean DropSense 16. Percent recovery was calculated based on the measured DNA concentration and the elution volume.
Figure 4: Monarch Mag PCR & DNA Cleanup Kit (5 µg) effectively removes enzymes compared to a leading supplier’s product
Monarch Mag PCR & DNA Cleanup Kit (5 µg) (NEB #T4130) effectively removes enzymes compared to a leading supplier’s product. DNA samples containing 0.8 units of Proteinase K (NEB #P8107) were purified using the Monarch Mag PCR & DNA Cleanup Kit (5 µg) and the leading supplier’s product, following manufacturer’s recommended protocol. Eluted DNA, along with control samples, was tested for residual Proteinase K activity by incubating with MBP5 (a Proteinase K substrate) in 1X NEBuffer™ 4 at 37 °C for 5 minutes. Reaction products were resolved on a Tris-Glycine polyacrylamide gel to assess protein integrity. A single, intact MBP5 band indicated complete removal of Proteinase K, whereas residual Proteinase K activity was evident by the appearance of multiple digestion fragments resembling the input (I) lane.
Monarch Mag PCR & DNA Cleanup Kit (5 µg) (NEB #T4130) workflow. Overview of workflow of the Monarch Mag PCR & DNA Cleanup Kit (5 µg) for manual or automated magnetic bead-based purification of DNA samples.
Figure 6: Monarch Mag PCR & DNA Cleanup Kit (5 µg) is scalable for various reaction volumes and DNA inputs
Monarch Mag PCR & DNA Cleanup Kit (5 µg) (NEB #T4130) is scalable for various reaction volumes and DNA inputs. The Monarch Mag PCR & DNA Cleanup Kit (5 µg) offers a flexible, high-performance solution for purifying a wide range of reaction volumes and DNA input amounts. While the kit includes materials optimized for standard reaction sizes, it also supports easy scaling up or down. Smaller- or higher-capacity workflows is possible using the provided supplemental protocol guidance.
This product is related to the following categories:
High Performance: Recover up to 5 μg of DNA with high yield and purity. Effectively remove primers, detergents, nucleotides, DMSO, betaine, and other low-molecular-weight reaction components.
Reliability: Consistent results for both recovery and purity.
Flexible Workflow: Compatible with both manual and automated processing.
Scalability: Easily adapted to different binding capacity requirements.
Broad Sample Compatibility: Purify a wide range of fragment sizes, from very small (< 50 bp) oligonucleotides to large (> 25 kb) DNA fragments, using the provided modified protocols.
Application Ready: Purified DNA is suitable for downstream molecular biology applications, including restriction digestion, ligation, amplification, sequencing, and in vitro transcription.
Application Features
Reaction Sample Types
PCR cleanup
DNA from PCR reactions can be purified after amplification to remove polymerases, primers, detergents, dNTPs, etc.
Enzymatic reaction cleanup
Restriction enzymes and modifying enzymes such as ligases, kinases, nucleases, phosphatases are efficiently removed, allowing for effective desalting and concentration of the DNA sample.
cDNA cleanup
DNA/RNA complexes can be purified post-reverse transcription/ amplification to enable removal of the RT and polymerase as well as nucleotides.
Labeling cleanup
Unincorporated radiolabeled or fluorescently labeled nucleotides can be removed from the DNA substrate.
Plasmid cleanup
Plasmid preps from unknown sources may contain inhibitors and unwanted contaminants. Purification and concentration can be easily achieved using this kit.
DNA Types
Oligonucleotide and ssDNA Purification
ssDNA oligonucleotides (≥ 12 nt) and dsDNA fragments (≥ 10 bp) can be purified using the supplemental protocol provided on the product webpage.
Genomic DNA, DNA size > 25 kb
gDNA and DNA size >25 kb can be purified using supplemental protocol provided on the product webpage.
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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