RNA Cleanup

Following 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.

 

 

Historically, RNA has been cleaned up in various ways, including phenol/chloroform extraction followed by ethanol precipitation, lithium chloride precipitation, or by using agarose gel electrophoresis. Silica-based spin columns have become a popular tool to clean up RNA. Spin column-based cleanup also offers a simple way to concentrate the RNA of interest at the same time by utilizing low elution volumes. NEB is proud to offer high performance, easy to use RNA cleanup kits in a variety of binding capacities for all your RNA workflows.

 

 

The Monarch Spin RNA Cleanup Kits provide a fast, simple silica spin column-based solution for RNA cleanup and concentration after any enzymatic reaction and after other purification methods. The Monarch Spin 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 can be purified with this kit; however, a modified protocol is available to enable the binding of RNA as small as 15 nt (including miRNAs).

 

Utilize Monarch RNA Cleanup solutions for a variety of applications

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

 

Optimize binding capacity and elution volume with Monarch Spin RNA Cleanup

Properties		Monarch Spin RNA Cleanup Kit (10 µg) NEB #T2030 S/L	Monarch Spin RNA Cleanup Kit (50 µg) NEB #T2040 S/L	Monarch Spin RNA Cleanup Kit (500 µg) NEB #T2050 S/L	Monarch Spin High-Capacity RNA Cleanup Kit (3 mg) NEB #T2060 S/L
	Maximum RNA Binding Capacity	Up to 10 µg	Up to 50 µg	Up to 500 µg	Up to 3 mg
	Format	Spin column
	Vacuum Compatibility	Yes			No
	Elution Volume	6–20 µl	20–50 µl	50–100 µl	250–1000 µl
	RNA Size Range	≥ 25 nt (≥ 15 nt with modified protocol)			≥ 25 nt
	Best for	Low-input RNA purification and concentration	Routine RNA purification	High-yield IVT and larger-scale RNA purification and concentration	Scaled-up IVT and mg-scale RNA purification
	Prep time	5 minutes spin and incubation		10–15 minutes spin and incubation time	25–30 minutes spin and incubation time

 

Purify Large Amounts of RNA from IVT Reactions with Monarch RNA Cleanup

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

 

Monarch RNA cleanup products provide fast and reliable purification of high-quality RNA after any enzymatic reaction.