Monarch® RNA Cleanup Kit (500 μg)

#T2050

Catalog # Size Price (£)
T2050L 100 PREPS £451.00
T2050S 10 PREPS £56.00

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.

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.

Properties & Usage

FAQs

Quality Control Assays

Certificate of Analysis

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 following link is to a PDF Safety Data Sheet (SDS) that applies to this product to help you use it safely.



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