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)|
|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|
- Can I purchase Monarch® buffers and columns separately?
- Are the columns in the Monarch RNA Cleanup Kits the same as those in the Monarch Total RNA Miniprep Kit (NEB #T2010)?
- Can I get better recovery with the Monarch RNA Cleanup Kits if I do a second elution with my eluent from the first elution?
- Can I use the Monarch RNA Cleanup Kit to cleanup up my DNase I-treated RNA?
- Can I use the Monarch RNA Cleanup Kits to cleanup my in vitro transcription (IVT) reaction?
- Can I use the Monarch RNA Cleanup Kits to cleanup RNA after a TRIzol®/chloroform extraction?
- Can I use the Monarch 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 RNA Cleanup Kit (NEB #T2040) compatible with the EnGen sgRNA Synthesis Kit, S. pyogenes?
- What factors affect my (A260/A230) when using the Monarch RNA Cleanup Kits?
- What is the composition of each buffer provided with the Monarch RNA Cleanup Kits?
- What is the maximum binding capacity of the Monarch RNA Cleanup Column provided with the Monarch RNA Cleanup Kit?
- What is the smallest volume of nuclease-free water that can be used for elution with the Monarch RNA Cleanup Columns?
- What size RNA can be purified with the Monarch RNA Cleanup Kit?
- Are the Monarch RNA Cleanup Kits compatible with Luna RT-qPCR reagents?
- Are the Monarch RNA Cleanup Kits compatible with NEBNext reagents for RNA library prep?
- My sample turned cloudy after adding the Monarch RNA Cleanup Binding Buffer and ethanol. Is this normal?
- Why do I need to incubate my column for 5 minutes with elution buffer (nuclease-free water)?
- Can I do an on-column DNase I treatment with the Monarch RNA Cleanup Columns?
- Monarch® RNA Cleanup Kit Protocol
- Purification of RNA from the Aqueous Phase Following TRIzol®/Chloroform Extraction using the Monarch® RNA Cleanup Kits
- Extraction of RNA from Agarose Gels using the Monarch® RNA Cleanup Kits
- Separation of Large and Small RNA into Fractions using the Monarch® RNA Cleanup Kits
- RNA Reaction Cleanup using the Monarch Total RNA Miniprep Kit (NEB #T2010)
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|
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Learn how to use the Monarch RNA Cleanup Kits to quickly and easily purify RNA from enzymatic reactions including in vitro transcription reactions, labeling, and DNase I treatment.
Optimize the results of your RNA cleanups with these quick and simple tips.
Learn about the sources of RNase contamination and how you can prevent them from disrupting your experiments.