Product Class: Kit

Monarch® Total RNA Miniprep Kit

Effective May 2, 2024, the Monarch gDNA Removal Columns are now tinted orange to help distinguish from other columns.

Product Introduction

Quickly and easily purify up to 100 µg of high quality total RNA from multiple sample types – just with this single, comprehensive kit.

  • For use with blood, cells and tissues, saliva, and buccal/nasopharyngeal swabs
  • Also works with tough to lyse samples (bacteria, yeast, plant)
  • Effectively purifies total RNA of all sizes, including small RNAs >20 nt
  • Efficient genomic DNA removal (column and DNase I-based)
  • Includes Proteinase K for processing of tissues and blood samples
  • Includes DNA/RNA Protection Reagent for sample preservation
  • Kit components and columns also available separately

Check out our Technical Note containing comprehensive insights into measuring and analyzing nucleic acids.

Catalog # Size Concentration
T2010S 50.0 preps

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Product Information

Description


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

monarch
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

monarch
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

monarch
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:
Total RNA Extraction & Purification Products,
RNA Extraction and Purification,
Nucleic Acid Purification Products,

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Product Notes

  1. 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.

Protocols, Manuals & Usage

Protocols

  1. Quick Protocol for Total RNA Extraction (NEB #T2010)
  2. Total RNA Purification from Cultured Mammalian Cells using the Monarch Total RNA Miniprep Kit (NEB #T2010)
  3. Total RNA Purification from Tissues and Leukocytes using the Monarch Total RNA Miniprep Kit (NEB #T2010)
  4. Total RNA Purification from Mammalian Whole Blood Samples using the Monarch Total RNA Miniprep Kit (NEB #T2010)
  5. Total RNA Purification from Tough-to-Lyse Samples (bacteria, yeast, plant, etc.) using the Monarch Total RNA Miniprep Kit (NEB #T2010)
  6. FFPE Tissue Deparaffinization and Subsequent RNA Purification Using the Monarch Total RNA Miniprep Kit  (NEB #T2010)
  7. Total RNA Purification from TRIzol®-extracted Samples using the Monarch Total RNA Miniprep Kit (NEB #T2010)
  8. Total RNA Purification from Samples in RNAlater® using the Monarch Total RNA Miniprep Kit (NEB #T2010)
  9. Total RNA Purification from Samples Stored in Monarch DNA/RNA Protection Reagent using the Monarch Total RNA Miniprep Kit (NEB #T2010)
  10. RNA Reaction Cleanup using the Monarch Total RNA Miniprep Kit (NEB #T2010)
  11. In-tube DNase I Treatment after Purification with the Monarch Total RNA Miniprep Kit (NEB #T2010)
  12. Fractionation of Small and Large RNA Using the Monarch Total RNA Miniprep Kit (NEB #T2010)
  13. RNA Extraction from Saliva, Buccal Swabs, and Nasopharyngeal Swabs using the Monarch Total RNA Miniprep Kit (NEB #T2010)
  14. Automation of RNA Extraction from Saliva Using the Monarch Total RNA Miniprep Kit

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

Usage & Guidelines

Application Notes

FAQs & Troubleshooting

FAQs

  1. Is the Monarch® Total RNA Miniprep Kit compatible with Luna® RT-qPCR Reagents?
  2. Is the Monarch® Total RNA Miniprep Kit compatible with NEBNext® reagents for RNA Library Prep?
  3. What is the lowest volume in which I can elute my RNA with the Monarch Total RNA Miniprep Kit (NEB #T2010)?
  4. Can the Monarch® Total RNA Miniprep Kit be used to purify RNA from yeast?
  5. Can the Monarch® Total RNA Miniprep Kit be used to purify RNA from bacteria?
  6. Can the Monarch® Total RNA Miniprep Kit be used to purify RNA from plants?
  7. How can I preserve my sample if I am not ready to extract RNA right after harvest?
  8. Can I purchase Monarch® buffers and columns separately?
  9. I am seeing genomic DNA in my preps when using the Monarch Total RNA Miniprep Kit. What could be the problem?
  10. Can I use the Monarch® Total RNA Miniprep Kit for RNA cleanup after enzymatic manipulations?
  11. Can I skip the on-column DNase I treatment step of the Monarch® Total RNA Miniprep Kit protocol?
  12. Can I skip the gDNA removal column step of the Monarch® Total RNA Miniprep Kit protocol?
  13. Can I use leftover Proteinase K and DNase I from the Monarch® Total RNA Miniprep Kit for other applications?
  14. Can I use NEB Proteinase K, Molecular Biology Grade (NEB #P8107S) in conjunction with the Monarch® Total RNA Miniprep Kit?
  15. Can the Monarch Total RNA Miniprep Kit (NEB #T2010) be used to purify RNA from saliva?
  16. Can the Monarch Total RNA Miniprep Kit NEB T2010 be used to purify RNA from buccal (cheek) swabs?
  17. Can the Monarch Total RNA Miniprep Kit (NEB #T2010) be used to purify RNA from plasma or serum?
  18. Can the Monarch Total RNA Miniprep Kit (NEB #T2010) be used to purify RNA from nucleated blood (e.g. bird, fish, reptile)?
  19. Can the Monarch Total RNA Miniprep Kit be used to purify RNA from drosophila?
  20. Can the Monarch Total RNA Miniprep Kit be used to purify RNA from zebrafish embryos/larvae?
  21. Can the genomic DNA captured on the Monarch gDNA Removal Column (NEB #T2017) be eluted and used in downstream applications?
  22. Can I lyse my adherent cells directly in their wells when using the Monarch Total RNA Miniprep Kit?
  23. How can I assess RNA integrity and purity?
  24. Can I use NEB DNase I (#M0303) with the Monarch Total RNA Miniprep Kit?
  25. I would like to purchase the contents of the Monarch Total RNA Miniprep Kit (NEB #T2010S) separately. Which products do I need?
  26. Can extraction of RNA with the Monarch kits be automated?
  27. Is live SARS-CoV-2 virus inactivated during extraction with the Monarch Total RNA Miniprep Kit?
  28. I noticed that in the Monarch Total RNA Miniprep Kit (NEB #T2010), the Monarch gDNA Removal Columns (NEB #T2017) are now tinted orange. Does this impact performance?
  29. I notice that I'm applying force to fit the column into a collection tube. Should I be doing this? What collection tubes are compatible with columns?
  30. Are the Monarch gDNA Removal Columns (NEB #T2017) and Monarch RNA Purification Columns (NEB #T2007) included with the Monarch Total RNA Miniprep Kit (NEB #T2010) compatible with the QIACube® System?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

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.

Specification 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 compatibility with QIACube has been updated.

Specifications

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]

Certificate Of Analysis

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]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

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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