Product Class: Other

Monarch® RNA Cleanup Columns (500 μg)

Product Introduction

The Monarch RNA Cleanup Columns (500 µg) are a component of the Monarch RNA Cleanup Kit (500 µg) (NEB #T2050) and can be used to purify up to 500 µg of RNA from enzymatic reactions. They are supplied with Monarch Collection Tubes II (NEB #T2018). RNA Cleanup Binding Buffer (NEB #T2041) and Monarch RNA Wash Buffer (NEB #T2042) are also available separately. 

  • For the cleanup of large amounts of RNA (up to 500 μg)
  • Ideal for purification after high yield in vitro transcription reactions
  • Elute in ≥ 50 µl for concentrated RNA
  • 70-100% RNA recovery
  • Unique column design prevents buffer carryover and elution of silica particulates
  • Purified RNA is ready for use in a wide variety of downstream applications

Learn more about Monarch DNA and RNA purification products

 

Catalog # Size Concentration
T2057L 100.0 columns

Product Information

Description

The Monarch RNA Cleanup Columns (500 µg) are a component of the Monarch RNA Cleanup Kit (500 µg) (NEB #T2050) and can be used to purify up to 500 µg of RNA from enzymatic reactions including high-yield RNA synthesis reactions. The columns are designed without the use of a retaining ring, ensuring no buffer retention and no carryover of contaminants. RNA can be eluted in as little as 50 μl for and is ready for use in a variety of downstream applications including RT-PCR, RNA Library Prep for NGS and transfection.

Monarch RNA Cleanup Columns are also available for 10 µg (NEB #T2037) and 50 µg (NEB #T2047) binding capacities.

 

Figure 1: The Monarch RNA Cleanup Kit (500 µg) is suitable for cleaning up large quantities (>250 µg) of RNA from in vitro transcription reactions

1. RNA transcripts of varying sizes (0.6-8 kb) were synthesized using the HiScribe®; T7 Quick High Yield RNA Synthesis Kit (NEB #E2050) using 1.5-1.8 µg of DNA template for two hours at 37°C. 40 µl of each in vitro transcription IVT) reaction was cleaned up using the Monarch RNA Cleanup Kit (500 µg, T2050) and eluted in 200 µl. RNA yields were calculated from the resulting A260, measured using a Nanodrop® spectrophotometer and ranged from 268-425 µg of RNA per IVT reaction.

2. RNA integrity (200 ng/lane) was assessed on a 1% agarose-TBE gel stained with SYBR Gold alongside the Millennium™ RNA Marker (Thermo Fisher Scientific).
Figure 2: RNA recovery is increased by incubating the column with the elution buffer (nuclease-free water) prior to the elution spin

rRNA (16S and 23S Ribosomal Standard from E.coli, Sigma) was purified using the Monarch RNA Cleanup Kit (500 µg, NEB #T2050). 100 µl of nuclease-free water was added to the column and incubated for either 0,1,3 or 5 minutes at room temperature before spinning to elute the RNA. The percent recovery of RNA was calculated from the resulting A260 as measured using a Trinean Dropsense 16. A five-minute incubation period produced the maximum yield.
This product is related to the following categories:
RNA Cleanup Products,
RNA Extraction and Purification,
Nucleic Acid Purification Products,

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Protocols, Manuals & Usage

Protocols

  1. Extraction of RNA from Agarose Gels using the Monarch® RNA Cleanup Kits
  2. Separation of Large and Small RNA into Fractions using the Monarch® RNA Cleanup Kits
  3. Monarch® RNA Cleanup Kit Protocol
  4. Download Quick Protocol Card Monarch RNA Cleanup Kit
  5. RNA Extraction from Cells Using the Monarch RNA Cleanup Kits

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. What is the maximum binding capacity of the Monarch RNA Cleanup Column provided with the Monarch RNA Cleanup Kit?
  2. What is the smallest volume of nuclease-free water that can be used for elution with the Monarch RNA Cleanup Columns?
  3. Can I get better recovery with the Monarch RNA Cleanup Kits if I do a second elution with my eluent from the first elution?
  4. What factors affect my (A260/A230) when using the Monarch RNA Cleanup Kits?
  5. What size RNA can be purified with the Monarch RNA Cleanup Kit?
  6. Can I use the Monarch RNA Cleanup Kit to cleanup up my DNase I-treated RNA?
  7. Can I use the Monarch RNA Cleanup Kits to purify RNA from agarose gels?
  8. Can I use the Monarch RNA Cleanup Kits to cleanup RNA after a TRIzol®/chloroform extraction?
  9. Can I use the Monarch RNA Cleanup Kits to cleanup my in vitro transcription (IVT) reaction?
  10. Are the Monarch RNA Cleanup Kits compatible with Luna RT-qPCR reagents?
  11. Are the Monarch RNA Cleanup Kits compatible with NEBNext reagents for RNA library prep?
  12. My sample turned cloudy after adding the Monarch RNA Cleanup Binding Buffer and ethanol. Is this normal?
  13. Why do I need to incubate my column for 5 minutes with elution buffer (nuclease-free water)?
  14. How can I assess RNA integrity and purity?
  15. Are the columns in the Monarch RNA Cleanup Kits the same as those in the Monarch Total RNA Miniprep Kit (NEB #T2010)?
  16. Can I purchase Monarch® buffers and columns separately?
  17. Can I do an on-column DNase I treatment with the Monarch RNA Cleanup Columns?
  18. Can the Monarch RNA Cleanup Kits (NEB #T2030, #T2040, #T2050) also be used to purify DNA?
  19. I would like to clean up  ~150 µg of RNA. Would it be better to use multiple 50 µg columns (NEB #T2040/#T2047) or a single 500 µg column (NEB #T2050/#T2057)?

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]

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 [email protected].

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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