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RNA Cleanup

Monarch RNA cleanup products provide fast and reliable purification of high-quality RNA after any enzymatic reaction.

Following RNA synthesis by in vitro transcription (IVT), unincorporated nucleotides, aborted transcripts, enzymes and buffer components should be removed before using the transcript for RNP formation or for microinjection. Removal of reactants is also beneficial following standard protocols such as RNA labeling, capping, Proteinase K treatment, and DNase I treatment. Sensitive workflows such as RNA-seq or RT-qPCR may also benefit from RNA cleanup prior to processing.

 

Workflow of RNA cleanup nucleic acid purification

 

Historically, RNA has been cleaned up in various ways, including phenol/chloroform extraction followed by ethanol precipitation, lithium chloride precipitation, or by using agarose gel electrophoresis. Silica-based spin columns have become a popular tool to clean up RNA. Spin column-based cleanup also offers a simple way to concentrate the RNA of interest at the same time by utilizing low elution volumes. NEB is proud to offer high performance, easy to use RNA cleanup kits in a variety of binding capacities for all your RNA workflows.

 

Four icons representing advantages of silica-based RNA cleanup

 

The Monarch Spin RNA Cleanup Kits provide a fast, simple silica spin column-based solution for RNA cleanup and concentration after any enzymatic reaction and after other purification methods. The Monarch Spin RNA Cleanup Kits are available in 3 different binding capacities: 10 μg (NEB #T2030), 50 μg (NEB #T2040) and 500 μg (NEB #T2050). Each kit contains unique columns, all designed to prevent buffer retention and ensure no carryover of contaminants, enabling low-volume elution of highly pure RNA (T2030: ≥ 6 μl, T2040: ≥ 20 μl and T2050: ≥ 50 μl). Following the standard protocol, RNA ≥ 25 nt can be purified with this kit; however, a modified protocol is available to enable the binding of RNA as small as 15 nt (including miRNAs).

 

Utilize Monarch RNA Cleanup solutions for a variety of applications

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

 

Optimize binding capacity and elution volume with Monarch Spin RNA Cleanup

Monarch Spin RNA Cleanup Kit

NEB #T2030 (10 µg)

NEB #T2040 (50 µg)

NEB #T2050 (500 µg)

Binding Capacity

10 μg

50 µg

500 µg

RNA Size Range

≥ 25 nt ( ≥ 15 nt with modified protocol)

Typical Recovery

70–100%

Elution Volume

6–20 µl

20–50 µl

50–100 µl

Purity

A260/280 > 1.8 and A260/230 > 1.8

Protocol Time

5 minutes of spin and incubation time

10–15 minutes of spin and incubation time

Common Downstream Applications

RT-PCR, RNA library prep for NGS, Small RNA library prep for NGS, RNA labeling

RT-PCR, RNA library prep for NGS, formation of RNP complexes for genome editing, microinjection, RNA labeling, transfection

RT-PCR, RNA library prep for NGS, RNA labeling, RNAi, microinjection, transfection

 

Purify Large Amounts of RNA from IVT Reactions with Monarch RNA Cleanup

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

Graph and gel showing cleaning large quantities 

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 Spin RNA Cleanup Kit (500 µg, NEB #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.

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RNA Cleanup
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Monarch StabiLyse DNA/RNA Buffer

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Monarch® Buffer BX

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Monarch® Buffer WX

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Monarch® Spin Columns S1A and Tubes

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Monarch® Spin Columns S2A and Tubes

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Monarch® Spin Columns S2B and Tubes

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Monarch® Spin RNA Cleanup Kit (10 μg)

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Monarch® Spin RNA Cleanup Kit (50 μg)

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Monarch® Spin RNA Cleanup Kit (500 μg)


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FAQs for RNA Cleanup
Protocols for RNA Cleanup
Application Notes for RNA Cleanup
Legal Information

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. All other trademarks are the property of their respective owners. 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.

 


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