The Monarch RNA Cleanup Kit (50 µg) rapidly and reliably purifies up to 50 μg of concentrated, high-quality RNA (> 25 nt) from enzymatic reactions including labeling, capping, in vitro transcription (IVT) and DNase I treatment. This kit utilizes a bind/wash/elute workflow with minimal incubation and spin times. Our unique column design ensures zero buffer retention and no carryover of contaminants, enabling elution of sample in volumes as low as 20 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 labeling. The protocol can also be modified to enable the purification of smaller RNA fragments (≥ 15 nts).
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 500 µg (NEB #T2050) binding capacities. Columns and buffers are also available separately for convenience.
Figure 1: Monarch RNA Cleanup Kit workflow
Specifications and Applications::
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SPECIFICATIONS
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RNA Sample Type
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Cleanup of previously purified RNA (TRIzol, phenol/ chloroform) and RNA from enzymatic reactions (in vitro transcription reactions, DNase I treatment)
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Binding Capacity
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50 µg
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RNA Size Range
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≥ 25 nt ( ≥ 15 nt with modified protocol)
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Typical Recovery
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70-100%
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Elution Volume
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20-50 µl
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Purity
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A260/280 > 1.8 and A260/230 > 1.8
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Protocol Time
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5 minutes of spin and incubation time
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Compatible Downstream Applications
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RT =-PCR, RNA library prep for NGS, Formation of ribonucleoprotein (RNP) complexes for genome editing studies, microinjection, RNA labeling, transfection
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APPLICATIONS
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RNA Cleanup and Concentration (including from the TRIzol aqueous phase)
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RNA purified by other methods can be further purified
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Enzymatic Reaction Cleanup
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Enzymes such as RNA polymerases, DNase I, Proteinase K and phosphatases are removed allowing efficient desalting
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In vitro Transcription Cleanup
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Enzymes and excess NTPs are removed to yield highly pure synthesized RNA
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RNA Gel Extraction
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Purification of RNA from agarose gels
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RNA Fractionation
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Fractionation of RNA into small and large RNA pools
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Figure 2: The Monarch RNA Cleanup Kit enables efficient recovery of RNA in as little as 20 µl
rRNA (50 µg of 16S and 23S Ribosomal Standard from E. coli, Sigma) was purified using the Monarch RNA Cleanup Kit (50 µg, NEB #2040) and eluted with nuclease-free water using the elution volumes indicated. The percent recovery of RNA was calculated from the resulting A260, as measured using a Trinean DropSense™ 16. Approximately 80% of RNA can be efficiently recovered in as little as 20 µl. Figure 3: The Monarch RNA Cleanup Kit (50 µg) produces sgRNA yields consistent with other competitor RNA cleanup kits and with lower residual NTP contamination
Six different sgRNA synthesis reactions from the EnGen® sgRNA Synthesis Kit, S. pyogenes (NEB #E3322) were cleaned up using either the Monarch RNA Cleanup Kit (50 µg, NEB #T2040) or a competitor kit (according to manufacturer's recommendations) and eluted with 50 µl of nuclease-free water. sgRNA yield was calculated from the resulting A260, as measured using a Trinean Dropsense™ 16. The Monarch Cleanup Kit produced sgRNA yields consistent with other commercially available RNA cleanup kits.
Following cleanup, residual nucleotides (NTPs) were measured by LC-MS and are reported as (percent area NTPs (rATP rCTP rGTP rUTP)/percent area sgRNA). The Monarch RNA Cleanup Kit consistently outperforms other commercially available RNA cleanup kits in the removal of residual NTPs from sgRNA synthesis reactions.
Properties & Usage
Protocols
Manuals
The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.
Quality Control Assays
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
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.
Legal and Disclaimers
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. 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.
New England Biolabs (NEB) is committed to practicing ethical science - we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.