Description
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.
Need automation support or a script? Submit a request using the Monarch Automation Support & Script Request form. Automation scripts are currently available for select platforms.
| 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.
Figure 5: Monarch Mag PCR & DNA Cleanup Kit (5 µg) workflow
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:
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Magnetic Bead-Based Nucleic Acid Purification,
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PCR & DNA Cleanup,
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Nucleic Acid Purification,
- This product can be used in the following applications:
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Nucleic Acid Purification