The Monarch Total RNA Miniprep Kit is a comprehensive solution for sample preservation, cell lysis, gDNA removal, and purification of total RNA from a wide variety of biological samples, including cultured cells, blood, and mammalian tissues. Additionally, tough-to-lyse samples, such as bacteria, yeast, and plant, can be processed with additional steps that enhance lysis. Cleanup of enzymatic reactions or purification of RNA from TRIzol® -extracted samples is also possible using this kit. Purified RNA has high quality metrics, including A260/280 and A260/230 ratios >1.8, high RIN scores, and minimal residual gDNA. Captured RNA ranges in size from full-length rRNAs down to intact miRNAs. Additionally, differential binding conditions allow selective capture or exclusion of the sub-200 nucleotide RNA pool that includes miRNA, 5S rRNA, and tRNA. Purified RNA is suitable for downstream applications, such as RT-qPCR, cDNA synthesis, RNA-seq, Northern blot analysis, etc.
Workflow for the Monarch Total RNA Miniprep Kit
Properties:
Binding Capacity:
100 µg RNA
RNA Size:
≥ 20 nt
Purity:
A260/280 typically ≥ 2.0, A260/230 usually ≥ 1.8
Tissue Input:
up to 20 mg for high yield tissues
up to 50 mg for low yield tissues
Cultured Mammalian Cells and Blood Cells (PBMC's, WBC's) Input:
up to 1 x 106
Mammalian Whole Blood Input:
up to 3 ml
Bacteria Input:
up to 1 x 109
Yeast Input:
up to 5 x 107
Plant Input:
up to 100 mg
Elution Volume:
30 - 100 μl
Yield:
varies depending on sample type
Compatible downstream applications:
RNA Library prep for NGS, RT-PCR, RT-qPCR, Northern blots
Tested Sample Types:
HeLa Cells
HEK 293 Cells
NIH 3T3 Cells
Human Blood
Rat Blood
PBMC's
Plasma
Serum
Nucleated Blood
Rat Liver
Rat Spleen
Rat Kidney
Rat Brain
Rat Muscle
Mouse Muscle
Mouse Heart
Mouse Kidney
Zebrafish Larvae
D. melanogaster
S. cerevisiae
E. coli
B. cereus
Tomato Leaf
Corn Leaf
Saliva
Buccal cells
Additional Data Figures:
Figure 1. 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).
Figure 2. 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).
Figure 3. 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.
This product is related to the following categories:
Monarch DNase I and Monarch Proteinase K can be stored at room temperature before reconstitution. After reconstitution, please store the enzymes at -20°C. We recommend aliquoting DNase I to minimize freeze-thaw cycles.
Cakar F, Zingl FG, Moisi M, Reidl J, Schild S. (2018) In vivo repressed genes of Vibrio cholerae reveal inverse requirements of an H+/Cl- transporter along the gastrointestinal passage. Proc Natl Acad Sci U S A; PubMedID: 29463743
Stephanie S. Lehman, Nicholas F. Norieab, Karin Aistleitnerb, Tina R. Clarkb, Cheryl A. Dooleyb, Vinod Nairc, Simran Jeet Kaura, M. Sayeedur Rahmana, Joseph J. Gillespiea, Abdu F. Azada, Ted Hackstadtb. (2018) The Rickettsial Ankyrin Repeat Protein 2 Is a Type IV Secreted Effector That Associates with the Endoplasmic Reticulum. mBio; PubMedID: 29946049
Quality, Safety & Legal
Quality Control Assays
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications & Change Notifications
Effective May 2, 2024, the Monarch gDNA Removal Columns are now tinted orange to help distinguish from other columns. The maximum input has been updated for cells.
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
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