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::
SPECIFICATIONS
RNA Sample Type
Cleanup of previously purified RNA (TRIzol, phenol/ chloroform) and RNA from enzymatic reactions (invitrotranscription 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 editingstudies, 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 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.
This product is related to the following categories:
RNA Cleanup Products,
RNA Reagents Products,
RNA Extraction & Purification Products,
Nucleic Acid Purification Products
This product can be used in the following applications:
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