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RNA Cleanup

As appreciation of the importance of RNA in biology continues to grow, the ability to quickly modify and manipulate RNA is in high demand. Accordingly, the need for rapid and reliable RNA cleanup methods have become essential. For example, after 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.

RNA can be cleaned up in various ways, including phenol/chlorform extraction followed by ethanol precipitation, lithium chloride precipitation, or by using agarose gel electrophoresis. More recently, silica-based spin columns have become a popular tool to clean up RNA. Additionally, spin column-based cleanup affords an easy way to concentrate the sample of interest at the same time, using low elution volumes. NEB is proud to offer a family of high performance and easy to use RNA cleanup kits for all your RNA workflows.

The Monarch RNA Cleanup Kits provide a fast and simple silica spin column-based solution for RNA cleanup and concentration after any enzymatic reaction (including in vitro transcription, DNase I treatment, capping and labeling) and after other purification methods such as phenol/chloroform extraction. The Monarch 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 is purified with this kit; however, a modified protocol is available to enable the binding of RNA as small as 15 nt (including miRNAs).




Monarch 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

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FAQs for RNA Cleanup
Protocols for RNA Cleanup
Application Notes for RNA Cleanup
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
Product Manual
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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.

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