RNA Extraction kit

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.

Feature

Reasons to choose the Monarch Total RNA Miniprep Kit

1
Purify high-quality RNA from multiple sample types and from low input amounts
2
Purify total RNA of all sizes, including RNA <200 nts
3
Generate RNA suitable for RT-qPCR and RT-PCR
4
Prepare high quality RNA-seq libraries

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 (NEB #T2010) 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).

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

Plasmid DNA encoding constitutively expressed GFP (pEGFP-C2) was prepared using either Monarch Plasmid Miniprep Kit or Qiagen QIAprep Spin Miniprep Kit. Four different cell lines (Cos-7, HEK293, HeLa, and Huh-7) were grown to 80-90% confluence and transfected with 100 ng of each plasmid, in complex with 0.3 μl Lipofectamine 2000, and 10 μl Opti-MEM. Five replicates for each cell type were performed using both DNA preps. GFP expressing cells were counted by flow cytometry 48 hrs post-transfection with a minimum of 2000 events collected per well. Average percentage of cells expressing GFP from all replicates is graphed and used as a measure of transfection efficiency.

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.

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).

Product Information

Total RNA Extraction & Purification

Monarch® RNA Purification Columns	Cat No.	SIze
Monarch® RNA Purification Columns	T2007L	100 columns
Monarch® RNA Lysis Buffer	Cat No.	SIze
Monarch® RNA Lysis Buffer	T2012L	100 ml
Monarch® RNA Priming Buffer	Cat No.	SIze
Monarch® RNA Priming Buffer	T2013L	56 ml
Monarch® RNA Wash Buffer	Cat No.	SIze
Monarch® RNA Wash Buffer	T2014L	50 ml

브랜드	NEBMonarch
개요	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.

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