Effective September 24, 2024, product name modified to Monarch® Spin RNA Cleanup Kit (50 μg). The component/product name of Monarch RNA Cleanup Columns has changed to Monarch Spin Columns S2A.
Effective March 25, 2024, component/product name changed to Monarch® Buffer BX.
Effective March 25, 2024, component/product name changed to Monarch® Buffer WX.
The Monarch Spin RNA Cleanup Kit (50 µg) enables fast and simple purification and concentration of up to 50 µg of RNA from enzymatic reactions.
Ideal for cleanup and concentration of RNA after enzymatic treatments including DNase I, Proteinase K, labeling, capping or in vitro transcription (IVT)
Efficiently purify RNA ≥ 25 nt (a simple modification enables purification of RNA ≥ 15 nt)
Elute in ≥ 20 µl for concentrated RNA
70-100% RNA recovery
Can be used to purify RNA from the aqueous phase following TRIzol® or similar extractions
Simplified workflow with a single wash buffer
Unique column design helps prevent buffer carryover and elution of silica particulates
Purified RNA is ready for use in a wide variety of downstream applications, including transfection
Check out our Technical Note containing comprehensive insights into measuring and analyzing nucleic acids.
The Monarch Spin 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 helps ensure 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 Spin 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 Spin RNA Cleanup Kit workflow
Specifications and Applications::
SPECIFICATIONS
RNA Sample Type
Cleanup of previously purified RNA (TRIzol, phenol/ chloroform) and RNA from enzymatic reactions (invitro
transcription reactions, DNase I treatment)
Binding Capacity
50 µg
RNA Size Range
≥ 25 nt ( ≥ 15 nt with modified protocol)
Typical Recovery
70–100%
Elution Volume
20–50 µl
Purity
A260/280 > 1.8 and A260/230 > 1.8
Protocol Time
5 minutes of spin and incubation time
Compatible Downstream Applications
RT =-PCR, RNA library prep for NGS, Formation of ribonucleoprotein (RNP) complexes for genome editing
studies, microinjection, RNA labeling, transfection
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
Figure 2: The Monarch Spin 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 Spin 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 Spin RNA Cleanup Kit (50 µg) produces sgRNA yields consistent with other suppliers' 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 Spin RNA Cleanup Kit (50 µg, NEB #T2040) or another supplier's 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 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 Spin RNA Cleanup Kit consistently outperforms other commercially available RNA cleanup kits in the removal of residual NTPs from sgRNA synthesis reactions.
This product is related to the following categories:
Check protocol to ensure correct buffer reconstitution, order of addition of buffers and ethanol, and proper handling of column flow-through and eluents.
Insufficient mixing of reagents
Ensure the ethanol is thoroughly mixed with RNA sample and RNA Cleanup Binding Buffer before applying the sample to the RNA Cleanup Column.
Incomplete elution during prep
Ensure the nuclease-free water used for elution is delivered directly to the center of the column so that the matrix is completely saturated. Larger elution volumes, multiple elutions, and longer incubation times can increase yield of RNA, but will dilute the sample and may increase processing times. For typical RNA samples, the recommended elution volumes and incubation times should be sufficient.
High degree of RNA secondary structure
Binding and elution of smaller RNAs (< 45 nt) can be affected by secondary structure of the RNA molecules. If poor yield of a small RNA is observed, we recommend diluting your sample with 2 volumes of ethanol instead of one volume in Step 2 of the protocol.
Purified RNA is Degraded
RNase contamination
In order to avoid RNase contamination during RNA cleanup, make sure to work on a clean lab bench, wear gloves and use disposable RNase-free pipet tips and microfuge tubes (not provided). Keep all kit components tightly sealed when not in use.
Improper storage of RNA
Purified RNA should be used immediately in downstream applications or stored at -70°C.
Low A260/230 Ratios
Residual guanidine salt carry-over
Ensure wash steps are carried out prior to eluting sample. Use care to ensure the tip of the column does not contact the flow-through. If unsure, repeat centrifugation. When reusing collection tubes, blot the rim of the tube on a Kimwipe prior to reattachment to the column to remove any residual wash buffer.
Low Performance of RNA in Downstream Steps
Salt and/or ethanol carry-over
Ethanol and salt remaining after the washes may inhibit downstream applications. Use care to ensure that the tip of the column does not come into contact with the flow-through. If in doubt, re-centrifuge for 1 minute to ensure traces of salt and ethanol are not carried over in the eluted RNA.
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.
Specifications & Change Notifications
Effective September 24, 2024, product name modified to Monarch® Spin RNA Cleanup Kit (50 μg). The component/product name of Monarch RNA Cleanup Columns has changed to Monarch Spin Columns S2A.
Effective March 25, 2024, component/product name changed to Monarch® Buffer BX. Product specifications have been updated.
Effective March 25, 2024, component/product name changed to Monarch® Buffer WX. Product specifications have been updated.
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 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]
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.
Based on your Freezer Program type, you are trying to add a product to your cart that is either not allowed or not allowed with the existing contents of your cart. Please review and update your order accordingly. If you have any questions, please contact Customer Support at freezers@neb.com or 1-800-632-5227 x 8.
You have been idle for more than 20 minutes, for your security you have been logged out. Please sign back in to continue your session.
Institution Changed
Your profile has been mapped to an Institution, please sign back for your profile updates to be completed.
Sign in to your NEB account
To save your cart and view previous orders, sign in to your NEB account. Adding products to your cart without being signed in will result in a loss of your cart when you do sign in or leave the site.
Powered by Bioz
