The Monarch Spin Plasmid Miniprep Kit offers a unique method for plasmid preparation and purification, enabling extraction of highly concentrated plasmid DNA from various bacterial strains, with yields up to 20 μg of high-quality DNA. This new version features upgraded spin columns precision engineered to uniquely allow for low elution, in as little as 30 μl, for highly concentrated DNA in downstream application such as sequencing, restriction digestion, transformation, transfection, and more. A colored buffer system helps indicate completion of key lysis and neutralization steps for an easy to follow workflow and shorter, faster protocols allow for reduced hands-on time and less spin time. Compared to leading suppliers, these kits offer significantly less plastic through uniquely designed columns and bottles with plastic reduction while still maintaining structural integrity for centrifugation or 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 for nucleic acid purification
Compatible methods
Centrifugation or vacuum manifold
Intended Usage
Plasmid extraction and preparation from various E.coli host strains and other microbial strains
Plasmid DNA Yield
Up to 20 μg*
Culture volume
1-5 ml, not to exceed 15 OD units
Column Binding Capacity
Up to 20 μg
Plasmid Size
Up to 25 kb
Elution Volume
≥ 30 μl
DNA Purity
A260/280 > 1.8 and A260/230 > 1.8
Protocol Time
9.5 - 12.5 minutes of spin and incubation time
Compatible Downstream Applications
Restriction digestion and other enzymatic manipulations, transformation, transfection, DNA sequencing, PCR, labeling, cell-free protein synthesis, etc.
* Yield depends on plasmid copy number, host strain, culture volume and growth conditions
FIGURE 1: Monarch Spin Plasmid Miniprep Kit consistently produces more concentrated plasmid DNA with equivalent or better yield and purity compared to the leading supplier
DNA concentration, yield and purity are higher with Monarch Spin Plasmid Miniprep Kit than leading supplier’s kit, across different plasmids. Preps were performed according to recommended protocols using ~1 ml (OD600 = 3) aliquot for pUC19 and pHT01 and ~2 ml (OD600 = 6) aliquot for Rluc and pXba of the same overnight cultures. Concentrations of plasmid were measured using a Trinean DropSense 16.
FIGURE 2: Plasmid DNA purified using Monarch Spin Plasmid Miniprep Kit are high-quality and suitable for downstream applications
Corresponding gel data shows purified plasmid DNA from Monarch Spin Plasmid Miniprep Kit works as well as leading supplier’s kit for downstream applications such as restriction digests. Gel data shows three replicates of plasmid purifications and subsequent restriction digests. Preps were performed according to recommended protocols using ~1 ml (OD600 = 3) aliquot for pUC19 and pHT01 and ~2 ml (OD600 = 6) aliquot for Rluc and pXba of the same overnight cultures. 1 μg of each prep was digested with XmnI (NEB #R0194) for pUC19, RLuc and pXba, and EcoRI (NEB #R0101) for pHT01 to linearize the plasmid and digests were resolved on 1.2% w/v TBE gel.
FIGURE 3: Monarch Spin Plasmid Miniprep Kit can elute in as low as 30 μl for high yield
Various elution volumes used in Monarch Spin Plasmid Miniprep Kit can generate high yield, highly concentrated DNA. 2 ml of NEB 10-beta Competent E. coli (NEB #C3019) transformed with pUC19 plasmid (NEB #N3041) was used. Monarch Buffer EY was used to elute the plasmid with the volumes indicated. Concentrations of plasmid were measured using a Trinean DropSense 16.
FIGURE 4: Features of the Monarch Spin Column S2D for Plasmid and Nucleic Acid Purification
NEB® Monarch’s unique column design and membrane assembly allows high-quality, highly-concentrated nucleic acid purification with low elution volume, for downstream applications. The column is designed and made with significantly less plastic for a reduced environmental impact.
FIGURE 5: Overview of the Monarch Spin Plasmid Miniprep Kit workflow
The Monarch Spin Plasmid Miniprep Kit features unique spin columns and optimized buffers based on alkaline lysis.
This product is related to the following categories:
High Performance: Achieve high yields (up to 20 μg) and high purity in the extraction and concentration of plasmid DNA without genomic DNA or RNA contamination.
High Concentration: Elute in as low as 30 μl, allowing for highly concentrated plasmid DNA.
Time Savings: 9.5- 12.5 minutes of spin and incubation time needed to complete the workflow.
Unique Design: The spin column features a unique design that enables elution in low volumes and minimizes buffer retention and contaminant carryover.
Optimized: The buffer system is optimized with color indicators to monitor the completion of each step.
Application Compatibility: Purified plasmid DNA is ready-to-use for restriction digestion and other enzymatic manipulations, transformation, transfection, DNA sequencing, PCR, labeling, cell-free protein synthesis, etc.
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]
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