RNA Cleanup
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.
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.
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
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APPLICATIONS |
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|---|---|
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RNA Cleanup and Concentration (including from the TRIzol aqueous phase) |
RNA purified by other methods can be further purified |
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Enzymatic Reaction Cleanup |
Enzymes such as RNA polymerases, DNase I, Proteinase K and phosphatases are removed allowing efficient desalting |
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In vitro Transcription Cleanup |
Enzymes and excess NTPs are removed to yield highly pure synthesized RNA |
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RNA Gel Extraction |
Purification of RNA from agarose gels |
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RNA Fractionation |
Fractionation of RNA into small and large RNA pools |
Optimize binding capacity and elution volume with Monarch Spin RNA Cleanup
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Monarch Spin RNA Cleanup Kit |
NEB #T2030 (10 µg) |
NEB #T2040 (50 µg) |
NEB #T2050 (500 µg) |
|---|---|---|---|
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Binding Capacity |
10 μg |
50 µg |
500 µg |
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RNA Size Range |
≥ 25 nt ( ≥ 15 nt with modified protocol) |
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Typical Recovery |
70–100% |
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Elution Volume |
6–20 µl |
20–50 µl |
50–100 µl |
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Purity |
A260/280 > 1.8 and A260/230 > 1.8 |
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Protocol Time |
5 minutes of spin and incubation time |
10–15 minutes of spin and incubation time |
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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
B. RNA integrity (200 ng/lane) was assessed on a 1% agarose-TBE gel stained with SYBR®Gold.
Choose Product:
Choose Type:
- Are the columns in the Monarch Spin RNA Cleanup Kits the same as those in the Monarch Total RNA Miniprep Kit (NEB #T2010)?
- Can I get better recovery with the Monarch Spin RNA Cleanup Kits if I do a second elution with my eluent from the first elution?
- Can I use the Monarch Spin RNA Cleanup Kit to cleanup up my DNase I-treated RNA?
- Can I use the Monarch Spin RNA Cleanup Kits to cleanup my in vitro transcription (IVT) reaction?
- Can I use the Monarch Spin RNA Cleanup Kits to cleanup RNA after a TRIzol®/chloroform extraction?
- Can I use the Monarch Spin RNA Cleanup Kits to purify RNA from agarose gels?
- Do you have a protocol for separating small and large RNAs into separate fractions?
- How can I assess RNA integrity and purity?
- Is the Monarch Spin RNA Cleanup Kit (NEB #T2040) compatible with the EnGen sgRNA Synthesis Kit, S. pyogenes?
- What factors affect my (A260/A230) when using the Monarch Spin RNA Cleanup Kits?
- What is the maximum binding capacity of the column for RNA Cleanup Column provided with the Monarch Spin RNA Cleanup Kit?
- What size RNA can be purified with the Monarch Spin RNA Cleanup Kit?
- Are the Monarch Spin RNA Cleanup Kits compatible with Luna RT-qPCR reagents?
- Are the Monarch Spin RNA Cleanup Kits compatible with NEBNext reagents for RNA library prep?
- Can I do an on column DNase I treatment with the columns for RNA Cleanup?
- Monarch® Spin RNA Cleanup Kit Protocol
- Purification of RNA from the Aqueous Phase Following TRIzol®/Chloroform Extraction using the Monarch® Spin RNA Cleanup Kits
- Extraction of RNA from Agarose Gels using the Monarch® Spin RNA Cleanup Kits
- Separation of Large and Small RNA into Fractions using the Monarch® Spin RNA Cleanup Kits
- RNA Reaction Cleanup using the Monarch Total RNA Miniprep Kit (NEB #T2010)
- Monarch RNA Purification Brochure
- RNA Metro Map Poster
- RNA Synthesis Brochure
- RNA Technical Guide
- Troubleshooting Guide for RNA Cleanup
- Guidelines for RNA Quantitation
- Guidelines for Working with RNA During RNA Cleanup
Brochures
Troubleshooting Guides
Usage Guidelines
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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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