New options now available: a magnetic-bead format with the Monarch Mag PCR & DNA Cleanup Kit (5 μg) (NEB #T4130), and a high-capacity spin‑column format for cleanup of up to 100 μg DNA with the Monarch Spin High-Capacity DNA Cleanup Kit (100 μg) (NEB #T1135).
New version of PCR and DNA cleanup kit, featuring upgraded spin columns precision-engineered for high performance
Elute in as little as 5 μl for highly concentrated DNA, with yields up to 5 μg
Prevent buffer retention and salt carry over with unique, optimized column design
No need to monitor pH
Protocol modification for oligonucleotide cleanup is provided, allowing purification of ssDNA, oligonucleotide and other small DNA fragments
Reduce hands-on time with faster protocols and less spin time
Significantly less plastic by design in columns and kit compared to leading suppliers
The Monarch Spin PCR & DNA Cleanup Kit (5 μg) offers an efficient method for purification and concentration up to 5 μg of high-quality, double-stranded and single-stranded DNA from enzymatic reactions such as PCR, restriction digestion, ligation, and reverse transcription. This new version features upgraded spin columns precision-engineered to uniquely allow for low elution, in as little as 5 μl, for highly concentrated DNA in downstream applications such as sequencing, restriction digestion, library construction, labeling and other enzymatic manipulations. The kit includes spin columns, collection tubes, buffers, and other reagents in compact and streamlined packaging for silica membrane-based PCR purification and DNA cleanup. Our optimized buffer system allows efficient binding and purification of the DNA, without a need to monitor pH. These kits contain uniquely designed columns and bottles to significantly reduce plastic usage compared to leading suppliers, while maintaining structural integrity for centrifugation, vacuum manifolds and other steps. The kit features Monarch spin columns that are manufactured using state of the art automation machinery for high quality and consistency.
Properties
Purification format
Spin-column
DNA sample
DNA from PCR and other enzymatic reactions (e.g., restriction digests, kinase reactions, ligations)
ssDNA or dsDNA oligonucleotides from enzymatic reactions can also be purified using the Oligonucleotide Cleanup Protocol
Typical recovery
70-90%
DNA Purity
A260/280 > 1.8 and A260/230 > 1.8
Binding Capacity
Up to 5 μg
Elution Volume
5-20 μl
DNA Size Range
Standard protocol: 50 bp – 25 kb
Protocol Time
5 minutes of spin and incubation time
Compatible Downstream Applications
Ligation, restriction digestion, labeling and other enzymatic manipulations, library construction and DNA sequencing
Figure 1: Monarch Spin PCR & DNA Cleanup Kit (5 μg) consistently produces more concentrated DNA, with equivalent or better DNA recovery and purity compared to the leading supplier
Using the Monarch Spin PCR & DNA Cleanup Kit (5 μg), DNA concentrations and recovery are equal to or higher than a leading suppliers' kits. Preps were performed according to recommended protocols and elution volumes using 5 μg of 3 kb linear DNA fragment as an input. Concentrations of DNA were measured using a Trinean DropSense 16 and percent recovery calculations are based on the eluted DNA concentration and elution volume used. Figure 2: DNA purified using Monarch® Spin PCR & DNA Cleanup Kit (5 μg) are high-quality and suitable for downstream applications
Use of the Monarch Spin PCR & DNA Cleanup Kit effectively cleans up DNA suitable for downstream applications, performing as well as a leading supplier’s kits.
A. TBE PAGE data demonstrating primer removal. A 2 kb fragment was spiked with two oligonucleotides (16-mer, 25-mer) to a final concentration of 1 μM. The mixture was purified as per recommended protocols. Equivalent fractions of the original mixture and the eluted DNA were resolved in a 20% TBE polyacrylamide gel at 100V for 1 hour and stained with SYBR Gold. M = Low Molecular Weight DNA Ladder (NEB #N3233).
B. Agarose gel data demonstrating downstream application by restriction digestion with a salt-sensitive restriction enzyme. Three replicates of purified DNA and subsequent restriction enzyme digest were performed. 4 μg of a 3 kb DNA fragment was purified and 1 μg was digested with DraIII-HF (NEB #R3510). Equivalent fractions of the input DNA and digestion reaction were resolved in a 1.2% TBE agarose gel. Figure 3: Monarch Spin PCR & DNA Cleanup Kit (5 μg) effectively cleans up oligonucleotide DNA using the oligonucleotide cleanup protocol.
Synthesized ssDNA (≥ 16 nt) and dsDNA (≥12 bp) oligonucleotides can be effectively purified and recovered using Monarch Spin PCR & DNA Cleanup Kit (5 μg). The provided oligonucleotide cleanup protocol was followed using 1 μg of oligonucleotides of varying lengths (8–25 nt/bp) as an input. DNA was eluted in 20 μl of Monarch Buffer EY. Concentrations of DNA were measured using a Trinean DropSense 16 and percent recovery calculations are based on the eluted DNA concentration and elution volume used. The minimum sized ss- and ds- oligonucleotides that can be used are marked with a star (*). Figure 4: Features of the Monarch Spin Column S1A
NEB® Monarch’s unique column design and membrane assembly allows high-quality DNA purification with low-elution volume, for highly-concentrated DNA for downstream applications. The column is designed and made with significantly less plastic for reduced environmental impact. Figure 5: Overview of the Monarch Spin PCR & DNA Cleanup Kit (5 μg) workflow
The Monarch Spin PCR & DNA Cleanup Kit (5 μg) uses the bind/wash/elute method and a unique spin column.
This product is related to the following categories:
High Performance: Achieve high yields (up to 5 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 5 μl for elution, allowing for highly concentrated DNA.
Powerful Flexibility – Purify small DNA fragments, including oligonucleotides, with this single kit, using the modified protocol provided.
Time Savings: Short protocol of only 5 minutes to complete the workflow of bind, wash, and elution steps, including minimal incubation and spin time.
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 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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