< Return to
Purify high-quality total RNA from cells, bloods, tissues and
other sample types using the Monarch Total RNA Miniprep Kit. This comprehensive kit includes genomic DNA removal columns, DNase I, Proteinase K and a stabilization/preservation reagent, all at a competitive price. Purified RNA ranges in size from full length RNA’s down to intact miRNAs and is ready for use in downstream applications including cDNA synthesis, RT-PCR, RT-qPCR and RNA-seq. Reasons to choose the Monarch Total RNA Miniprep Kit
Purify high-quality RNA from multiple sample types and low input amounts
Monarch-purified RNA is high-quality and compatible with a wide variety of downstream applications
Total RNA from a broad array of sample types was purified using the Monarch Total RNA Miniprep Kit ( NEB #T2010). Aliquots were run on an Agilent® Bioanalyzer ® 2100 using the Nano or Pico 6000 RNA chip (S. cerevisiae RNA was run using a plant Nano assay). RIN values and O.D. ratios confirm the overall integrity and purity of the RNA. To demonstrate compatibility with downstream applications, samples were subsequently used for RT-PCR (A) using Protoscript ® II Reverse Transcriptase ( NEB #M0368)/LongAmp ® Taq DNA Polymerase ( NEB #M0323), NGS library prep (B) using NEBNext ® Ultra™ II RNA Library Prep Kit ( NEB #E7760) and RT-qPCR (C) using Luna qPCR Reagents ( NEB #E3005).
The Monarch Total RNA Miniprep Kit can generate high quality RNA from as few as 100 HeLa cells
Total RNA was isolated using the Monarch Total RNA Miniprep Kit ( ) from varying amounts of HeLa cells over 5 orders of magnitude and eluted in 100 µl of nuclease-free water.Samples were analyzed on a Bioanalyzer Pico chip, with RIN values and total yields shown below the lanes (A). Electropherograms are included as a reference (B). NEB #T2010
Purify total RNA of all sizes, including RNA <200 nts
The Monarch Total RNA Miniprep Kit successfully purifies small RNAs below 200 nucleotides, enabling a more faithful representation of the total RNA pool
RNA preps were performed on HEK293 cells, mouse heart, rat brain, or rat spleen using the Monarch Total RNA Miniprep Kit (NEB #T2010) and the RNeasy Mini Kit from Qiagen. Equivalent amounts were resolved on a Bioanalyzer 2100 using the Small RNA chip. Monarch-purified RNA contains significantly more RNA in the sub-200 nucleotide pool.
Generate RNA suitable for RT-qPCR and RT-PCR
RT-qPCR on Monarch-purified RNA generates high-quality qPCR curves demonstrating accurate quantitation across a wide variety of sample types
Monarch-purified RNA from human whole blood, rat kidney, tomato leaf, and the yeast S.cerevisiae was diluted to produce a five-log range of input template concentrations. Primers targeting GAPDH variants (tomato, yeast), PGK (rat), or SMG1 (human blood) were used for RT-qPCR assays, assembled as directed using Luna RT-qPCR reagents (NEB #E3005) and cycled on a Bio-Rad CFX384.
View additional RT-qPCR and RT-PCR data
Prepare high quality RNA-seq libraries
Monarch-purified RNA can be used to prepare high quality RNA-seq libraries for gene expression analysis
Transcript levels in Universal Human Reference RNA (UHRR, Agilent) are compared before and after re-purification using either Qiagen RNeasy or the Monarch Total RNA Miniprep Kit. Strong correlation with untreated UHRR is observed for both methods (Pearson R > 0.99 for both samples). All samples display consistent end-to-end coverage of transcripts indicating an absence of detectable degradation during purification.
Poly-A selected RNA was selected from 100 ng of untreated, Qiagen and Monarch samples using the NEBNext Poly(A) mRNA Magnetic Isolation Module. RNA-seq libraries were then prepared using the NEBNext Ultra II Directional RNA Library Prep Kit before sequencing on an Illumina® Miseq® instrument (2 x150). 1.6M reads were randomly sampled from each library and adapter trimmed (Seqprep v1.1). Levels of all Gencode v26 transcripts were assessed using Salmon (0.4) and plotted above (panel A). Average 5´→3´ Coverage of Gencode v26 transcripts (assessed by Picard's CollectRnaSeqMetrics 1.56 after mapping to the GRCh38 reference genome with Hisat v2.0.3 and marking duplicates with Picard's MarkDuplicates 1.56) is shown below (panel B).