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Monarch® RNA Cleanup Kit (500 µg)

Catalog # Concentration Size List Price Quantity Your Price
T2050L 100 preps $741.00
$666.90
T2050S 10 preps $97.00
$87.30
Catalog # Size List Price Your Price
T2050L 100 preps $741.00
$666.90
T2050S 10 preps $97.00
$87.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

The Monarch RNA Cleanup Kit (500 µg) enables fast and simple purification and concentration of up to 500 µg of RNA from in vitro transcription (IVT) and other enzymatic reactions.

  • Ideal for purification of synthesized RNA following high-yield in vitro transcription reactions (alternative to MEGAclear™)
  • Optimized for the cleanup of RNA after enzymatic treatments including DNase I, Proteinase K, labeling and capping
  • Efficiently purify RNA ≥ 25 nt (a simple modification enables purification of RNA ≥ 15 nt)
  • Elute in ≥ 50 µl for concentrated RNA 70-100% RNA recovery
  • Unique column design prevents buffer carryover and elution of silica particulates
  • Simplified workflow with a single wash buffer
  • Columns and buffers are available separately
  • Purified RNA is ready for use in a wide variety of downstream applications, including microinjection and transfection.

Check out our Technical Note containing comprehensive insights into measuring and analyzing nucleic acids.

Featured Videos
View Video Library
  • Monarch_MiniprepProtocol_VideoThumb
    Monarch® RNA Cleanup Protocol
  • Monarch_MiniprepProtocol_VideoThumb
    Tips for successful RNA Cleanup using the Monarch® RNA Cleanup Kits
  • Monarch_MiniprepProtocol_VideoThumb
    Avoiding RNase Contamination
  • Monarch_MiniprepProtocol_VideoThumb
    How to recycle your Monarch® Kit components

The Monarch RNA Cleanup Kit (500 µg) reliably purifies up to 500 μg of concentrated, high-quality RNA (> 25 nt) from enzymatic reactions and in vitro transcription (IVT) reactions. The kit utilizes a bind/wash/elute workflow with minimal incubation and spin times. The columns ensure zero buffer retention and no carryover of contaminants, enabling elution of sample in volumes as low as 50 μl. Unwanted NTPs and short RNA fragments are removed, ensuring highly pure RNA transcripts following IVT/RNA synthesis. Eluted RNA is ready for use in a variety of downstream applications including RT-PCR, RNA Library Prep for NGS and RNA labelling. The protocol can also be modified to enable the purification of smaller RNA fragments (≥ 15 nt).

Designed with sustainability in mind, Monarch kits use significantly less plastic and responsibly-sourced, recyclable packaging.

Monarch RNA Cleanup Kits are also available for 10 µg (NEB #T2030) and 50 µg (NEB #T2040) binding capacities. Columns and buffers are also available separately for convenience.

 

Figure 1: Monarch RNA Cleanup Kit workflow

 

Specifications and Applications:

SPECIFICATIONS
RNA Sample Type Cleanup of RNA from large-scale in vitro transcription reactions
Binding Capacity 500 µg
RNA Size Range ≥ 25 nt ( ≥ 15 nt with modified protocol)
Typical Recovery 70–100%
Elution Volume 50–100 µl
Purity A260/280 > 1.8 and A260/230 > 1.8
Protocol Time 10–15 minutes of spin and incubation time
Compatible Downstream Applications RNA Labeling, RNAi, Microinjections, RT-PCR, RNA library prep for NGS, transfection

APPLICATIONS
RNA Cleanup and Concentration (including from the TRIzol aqueous phase) RNA purified by other methods can be further purified
Enzymatic Reaction Cleanup Enzymes such as RNA polymerases, DNase I, Proteinase K and phosphatases are removed allowing efficient desalting
In vitro Transcription Cleanup Enzymes and excess NTPs are removed to yield highly pure synthesized RNA
RNA Gel Extraction Purification of RNA from agarose gels
RNA Fractionation Fractionation of RNA into small and large RNA pools

 

Figure 2: The Monarch RNA Cleanup Kit (500 µg) is suitable for cleaning up large quantities (>250 µg) of RNA from in vitro transcription reactions

A. 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.

B. RNA integrity (200 ng/lane) was assessed on a 1% agarose-TBE gel stained with SYBR® Gold.
Figure 3: The Monarch RNA Cleanup Kit (500 µg) cleans up large-scale in vitro transcription reactions and generates yields consistent with other large-scale cleanup kits

0.3 and 1.8 kb fragments were in vitro transcribed at 37°C (overnight and 2 hours, respectively) using the HiScribe®; T7 Quick High Yield RNA Synthesis Kit (NEB #E2050). Following DNase I treatment (4 U DNase I, 37°C, 15 minutes), transcription reactions were pooled and 200 µl cleaned up using either the NEB Monarch RNA Cleanup Kit (500 µg) or the MEGAclear Transcription Clean-Up Kit (Thermo Fisher Scientific). In vitro transcribed RNA was eluted twice with 100 µl of nuclease-free water following a 5-minute on-column incubation (room temperature for Monarch and 65°C for MEGAclear. Recovery of the synthesized RNA transcript was calculated from the resulting A260 using a Trinean Dropsense™ 16. The Monarch RNA Cleanup Kit (500 µg) produces similar RNA yields as the MEGAclear Kit for large-scale in vitro transcription reactions.  
Figure 4: 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.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Monarch® RNA Cleanup Columns (500 μg) T2057-21 25 1 x 10 columns
  Monarch® Collection Tubes II T2018-21 25 1 x 10 tubes
  Monarch® Buffer BX T2041-21 25 1 x 3 ml
  Monarch® Buffer WX T2042-21 25 1 x 2.5 ml
  Nuclease-free Water B1500-21 25 1 x 2.5 ml
  Monarch® RNA Cleanup Columns (500 μg) T2057-1 25 2 x 50 columns
  Monarch® Collection Tubes II T2018-1 25 2 x 50 tubes
  Monarch® Buffer BX T2041-2 25 1 x 40 ml
  Monarch® Buffer WX T2042-2 25 2 x 20 ml
  Nuclease-free Water B1500-2 25 1 x 25 ml
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

FAQs
Troubleshooting
Quality Control Assay
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.
Change Notifications

Effective March 25, 2024, component/product name changed to Monarch® Buffer BX. Product specifications have been updated.

Effective March 25, 2024, component/product name changed to Monarch® Buffer WX. Product specifications have been updated.

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 Data Sheets
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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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