Monarch RNA Cleanup Kits – New England Biolabs GmbH

Monarch Kits for RNA Purification and Concentration

The Monarch RNA Cleanup Kits provide a fast and simple silica spin column-based solution for RNA purification and concentration after any enzymatic reaction (including in vitro transcription, DNase I treatment, capping and labeling) and after other isolation methods such as phenol/chloroform extraction.

Video: RNA Purification Protocol

The Monarch RNA Cleanup Kits are available in 3 different binding capacities:

for 10 μg (NEB #T2030)
for 50 μg (NEB #T2040)
and for 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. 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).

Reasons to choose Monarch for RNA Cleanup:

Choose from 3 different binding capacities and elution volumes
Quickly and easily purify large quantities of high-quality RNA from in vitro transcription (IVT) reactions
Efficiently remove unincorporated nucleotides from your RNA sample

Video: RNA Purification Tips

Typical 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

Customer feedback

„Sample Type: Yeast (fresh mycelium) (compared to: Qiagen RNeasy Mini Kit) “Better yield and quality; easy protocol and good and consistent results among samples.”“

Gisele, Research Associate, Mississippi State University

„Sample Type: Gram Positive Bacteria (compared to: Qiagen RNeasy Mini) “I've been using the RNeasy kit for some time, but I've been unable to have much luck. It seems geared toward eukaryotes and I've been working with Gram Positive prokaryotes… I tried the NEB `{`Total`}` RNA Miniprep Kit and it was much more straightforward. I also got excellent results the first time I used it.”“

Kyle, Postdoctoral Researcher, University of CA, Berkeley

„Sample Type: CD34 Cells “I repeated the RNA extraction using `{`NEB’s Monarch Total RNA Miniprep Kit`}`. The quality of RNAs were checked using Tape Station. And all four samples had RIN between 8.3 to 9.3, so they are all good…Your RNA extraction kit is easy to use and saves time.”“

Researcher, UCL Great Ormond Street Institute of Child Health

„Sample Type: Oat Protoplast Cells (compared to Qiagen RNeasy Plant Mini Kit) “I compared the yield of RNA extracted and the purity of the RNA based on spectrometer (Nanodrop). When I compared to my usual RNeasy kit from Qiagen to the Monarch sample, I obtain 5 times more RNA from my sample as well as higher purity of my sample with the Monarch RNA mini-prep.”“

Researcher, Iowa State University

„Sample Type: Mouse Heart (compared to: Thermo Fisher Scientific GeneJet RNA Isolation Kit) ``I liked the consistency, ease of use, and the fact that everything you need (e.g. DNase, Proteinase K) comes in the kit. Both the concentration and quality of the isolated RNA from the Monarch Kit were slightly better than the GeneJet kit. The bigger difference was in the consistency of the RNA isolations between samples--it was much better with the Monarch Kit.``“

Natalie, PhD. Student, Winnipeg, CA

„Sample Type: RNA from Virus in cell culture supernatant (compared to: TRIzol extraction) “I’ve had a go extracting RNA from a couple of different samples using the Monarch kit, in parallel with Trizol extractions from duplicate samples. The initial results, nanodrop analysis of the extracted RNA, were very positive.”“

Jessica Purcell, Pirbright Institute, United Kingdom

Flexibility in Binding Capacity and Elution Volume

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)
Typical Recovery:	70–100%
Elution Volume:	6–20 µl	20–50 µl	50–100 µl
Purity:	A_260/280 > 1.8 and A_260/230 > 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

Purify Large Amounts of RNA from IVT Reactions

rRNA (10, 50 or 500 µg, respectively of 16S and 23S Ribosomal Standard from E. coli, Sigma) was purified using a Monarch RNA Cleanup Kit (10 µg, NEB #T2030) (50 µg, NEB #T2040) (500 µg, NEB #T2050). Nuclease-free water was used to elute the RNA. The percent recovery of the RNA was calculated from the resulting A260 as measured using a Trinean DropSense 16. ~80% of RNA can be efficiently recovered in 6 µl from the Monarch RNA Cleanup Kit (10 µg, NEB #T2030), 20 µl from the Monarch RNA Cleanup Kit (50 µg, NEB #T2040), and 50 µl from the Monarch RNA Cleanup Kit (500 µg, NEB #T2050).

The Monarch RNA Cleanup Kit (500 µg) is suitable for cleaning up large quantities (>250 µg) of RNA from in vitro transcription reactions

1. RNA transcripts of varying sizes (0.6-8 kb) were synthesized using the HiScribe™ T7 Quick High Yield RNA Synthesis Kit (NEB #E2050) using 1.5-1.8 µg of DNA template for two hours at 37°C. 40 µl of each in vitro transcription IVT) reaction was cleaned up using the Monarch RNA Cleanup Kit (500 µg, T2050) and eluted in 200 µl. RNA yields were calculated from the resulting A260, measured using a Nanodrop® spectrophotometer and ranged from 268-425 µg of RNA per IVT reaction.

2. RNA integrity (200 ng/lane) was assessed on a 1% agarose-TBE gel stained with SYBR® Gold.

T2050_Fig2_MonarchRNACleanup_largeQuantities

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.

Available RNA Purification Kits:

PRODUCT	Prod. #	SIZE
Monarch RNA Cleanup Kit (10 µg)	T2030S	10 preps
Monarch RNA Cleanup Kit (10 µg)	T2030L	100 preps
Monarch RNA Cleanup Kit (50 µg)	T2040S	10 preps
Monarch RNA Cleanup Kit (50 µg)	T2040L	100 preps
Monarch RNA Cleanup Kit (500 µg)	T2050S	10 preps
Monarch RNA Cleanup Kit (500 µg)	T2050L	100 preps

As of: 01.01.2024

Also available:

PRODUCT	Prod.Nr.:	SIZE
Monarch RNA Cleanup Columns (10 µg)	T2037L	100 preps
Monarch RNA Cleanup Binding Buffer	T2041L	80 ml
Monarch RNA Cleanup Wash Buffer	T2042L	40 ml
Monarch RNA Cleanup Columns (50 µg)	T2047L	100 preps
Monarch RNA Cleanup Columns (500 µg)	T2057L	100 preps