Product Class: Kit

Monarch® Spin Plasmid Miniprep Kit


Catalog #T1110

Product Introduction

  • A new version of plasmid miniprep kit, featuring upgraded spin columns precision engineered for high performance
  • Elute in as little as 30 μl for highly concentrated DNA, with yields up to 20 μg
  • Prevent buffer retention and salt carry over with unique, optimized column design
  • Easy to follow workflow with colored buffer system to indicate completion of key steps
  • Reduce hands-on time with faster protocols and less spin time
  • Significantly less plastic by design in columns and kit compared to leading suppliers

T1110 Box

Product Information

Description

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

T1110 Concentration

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

T1110 Quality

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

T1110 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 

T1110 Column Design

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

T1110 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:
Plasmid Purification Products,
Nucleic Acid Purification Products,
This product can be used in the following applications:
Nucleic Acid Purification

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • T1110S     25    
      Monarch® Spin Columns S2D T1117-1 25 1 x 50 columns Not Applicable
      Monarch® Spin Collection Tubes T2118-1 25 1 x 50 tubes Not Applicable
      Monarch® Buffer B1 T1111-1 25 1 x 12 ml Not Applicable
      Monarch® Buffer B2 T1112-1 25 1 x 11 ml Not Applicable
      Monarch® Buffer B3 T1113-1 25 1 x 22 ml Not Applicable
      Monarch® Buffer BZ T1114-1 25 1 x 8.4 ml Not Applicable
      Monarch® Buffer WZ T1115-1 25 1 x 5 ml Not Applicable
      Monarch® Buffer EY T1116-1 25 1 x 7 ml Not Applicable
      Monarch® RNase A T3018-1 -20 1 x 0.17 ml 20 mg/ml
  • T1110L     25    
      Monarch® Spin Columns S2D T1117-1 25 5 x 50 columns Not Applicable
      Monarch® Spin Collection Tubes T2118-1 25 5 x 50 tubes Not Applicable
      Monarch® Buffer B1 T1111-2 25 1 x 57 ml Not Applicable
      Monarch® Buffer B2 T1112-2 25 2 x 27 ml Not Applicable
      Monarch® Buffer B3 T1113-2 25 1 x 110 ml Not Applicable
      Monarch® Buffer BZ T1114-3 25 1 x 42 ml Not Applicable
      Monarch® Buffer WZ T1115-3 25 1 x 26 ml Not Applicable
      Monarch® Buffer EY T1116-2 25 1 x 25 ml Not Applicable
      Monarch® RNase A T3018-2 -20 1 x 0.5 ml 20 mg/ml

Properties & Usage

Features

  • 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.

Protocols, Manuals & Usage

Protocols

  1. Plasmid Miniprep Protocol using Centrifugation (NEB #T1110)
  2. Plasmid Miniprep Protocol using a Vacuum Manifold (NEB #T1110)
  3. Buffer Preparation Guidance (NEB #T1110)

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

Usage & Guidelines

Application Notes

FAQs & Troubleshooting

FAQs

  1. Can I purchase Monarch columns and collection tubes separately?
  2. Can I use water to elute the DNA when using Monarch Kits?
  3. For the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110), which culture media do you recommended for growing E.coli?
  4. Which E.coli strain(s) are recommended for use with the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110)?
  5. What culture conditions do you recommended for optimal performance with the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110)?
  6. How can I prevent RNA contamination in DNA samples when using the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110)?
  7. What is the smallest volume of elution buffer that can be used with Monarch® Spin Plasmid Miniprep Kit (NEB #T1110)?
  8. What is the composition of each buffer provided with the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110)?
  9. When using the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110), why might I observe a smear above the plasmid band on an agarose gel?
  10. When using the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110), what would explain additional bands I see on the gel?
  11. What factors affect the recovery of plasmid DNA when using the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110)?
  12. What factors affect my (A260/280) after performing a plasmid miniprep using the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110)?
  13. What factors affect my (A260/230) after performing a plasmid miniprep using the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110)?
  14. What is the maximum binding capacity of the Monarch Plasmid Miniprep Column?
  15. What should I do differently when using a low copy plasmid?
  16. Do the dyes in the buffer of the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110) affect downstream DNA applications?
  17. Can I use DNA isolated from the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110) for transfection?
  18. Are Monarch® Spin Columns S2D (NEB #T1117) compatible with vacuum manifolds?
  19. Can the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110) guarantee endotoxin-free plasmid DNA recovery?
  20. Why might low yields of plasmid DNA or high levels of contaminating genomic DNA (gDNA) occur when using 5 ml of cell culture for plasmid isolation with the Monarch® Spin Plasmid Miniprep Kit (NEB #T1110), and how can this be improved?
  21. Can components from the older Monarch® Plasmid Miniprep Kit (NEB #T1010) be used interchangeably with those from the new version of the kit (Monarch Spin Plasmid Miniprep Kit, NEB #T1110)?
  22. Before use, how should I prepare the buffers? Do I need to add isopropanol and/or ethanol to the buffers prior to use?
  23. I notice that I'm applying force to fit the column into a collection tube. Should I be doing this? What collection tubes are compatible with columns?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

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

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]

Certificate Of Analysis

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]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

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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