The Monarch RNA Cleanup Columns (500 µg) are a component of the Monarch RNA Cleanup Kit (500 µg) (NEB #T2050) and can be used to purify up to 500 µg of RNA from enzymatic reactions including high-yield RNA synthesis reactions. The columns are designed without the use of a retaining ring, ensuring no buffer retention and no carryover of contaminants. RNA can be eluted in as little as 50 μl for and is ready for use in a variety of downstream applications including RT-PCR, RNA Library Prep for NGS and transfection.
Monarch RNA Cleanup Columns are also available for 10 µg (NEB #T2037) and 50 µg (NEB #T2047) binding capacities.
Figure 1: 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 alongside the Millennium™ RNA Marker (Thermo Fisher Scientific).Figure 2: RNA recovery is increased by incubating the column with the elution buffer (nuclease-free water) prior to the elution spin
rRNA (16S and 23S Ribosomal Standard from E.coli, Sigma) was purified using the Monarch RNA Cleanup Kit (500 µg, NEB #T2050). 100 µl of nuclease-free water was added to the column and incubated for either 0,1,3 or 5 minutes at room temperature before spinning to elute the RNA. The percent recovery of RNA was calculated from the resulting A260 as measured using a Trinean Dropsense 16. A five-minute incubation period produced the maximum yield.
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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