Monarch^® Total RNA Miniprep Kit

Product Information

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 A_260/280 and A_260/230 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

 

Figure 2. The Monarch Total RNA Miniprep Kit can generate high quality RNA from as few as 100 HeLa cells

 

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

 

This product is related to the following categories:

Total RNA Extraction & Purification Products,

RNA Extraction and Purification,

Nucleic Acid Purification

• Kit Components

  The following reagents are supplied with this product:

  NEB https://www.neb.com/en/#	Component Name	Component https://www.neb.com/en/#	Stored at (°C)	Amount	Concentration
  T2010S     25       Monarch® gDNA Removal Columns T2017-1 25 1 x 50 columns Not Applicable   Monarch® RNA Purification Columns T2007-1 25 1 x 50 columns Not Applicable   Monarch® Spin Collection Tubes T2118-1 25 2 x 50 tubes Not Applicable   Monarch® DNA/RNA Protection Reagent T2011-1 25 1 x 28 ml Not Applicable   Monarch® RNA Lysis Buffer T2012-1 25 1 x 50 ml Not Applicable   Monarch® Proteinase K Reaction Buffer T2003-1 25 1 x 4 ml Not Applicable   Monarch® DNase I Reaction Buffer T2005-1 25 1 x 4 ml Not Applicable   Monarch® RNA Priming Buffer T2013-1 25 1 x 28 ml Not Applicable   Monarch® RNA Wash Buffer T2014-1 25 1 x 25 ml Not Applicable   Monarch® Nuclease-free Water T2006-1 25 1 x 13 ml Not Applicable   Monarch® Proteinase K T2001-1 25 1 x 20 mg 20 mg/ml   Monarch® Proteinase K Resuspension Buffer T2002-1 25 1 x 1.5 ml Not Applicable   Monarch® DNase I T2004-1 25 1 x 275 units 1 units/μl

• Related Products

  Companion Products

  • Luna^® Universal One-Step RT-qPCR Kit
  • Luna^® Universal Probe One-Step RT-qPCR Kit
  • ProtoScript® II First Strand cDNA Synthesis Kit
  • NEBNext^® Ultra^™ II RNA Library Prep Kit for Illumina^®
  • NEBNext^® Ultra^™ II Directional RNA Library Prep Kit for Illumina^®
  • NEBNext^® Ultra^™ II Directional RNA Library Prep with Sample Purification Beads
  • NEBNext® Poly(A) mRNA Magnetic Isolation Module
  • ProtoScript® II Reverse Transcriptase

  Materials Sold Separately

  • Nuclease-free Water
  • Monarch^® DNA/RNA Protection Reagent
  • Monarch® RNA Purification Columns
  • Monarch^® gDNA Removal Columns
  • Monarch^® RNA Lysis Buffer
  • Monarch^® RNA Priming Buffer
  • Monarch^® RNA Wash Buffer
  • Monarch^® Total RNA Miniprep Enzyme Pack
  • Monarch^® Collection Tubes II

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

  • manualT2010

• Usage Guidelines

  • General Guidelines for Successful RNA Purification Using the Monarch Total RNA Miniprep Kit
  • Guidance on Choosing Sample Input Amounts when Using the Monarch Total RNA Miniprep Kit
  • Guidelines for Purification of RNA from Cultured Mammalian Cells
  • Guidelines for RNA Purification from Bacteria
  • Guidelines for RNA Purification from Leukocytes (WBCs, PBMCs)
  • Guidelines for RNA Purification from Plants
  • Guidelines for RNA Purification from Tissues 
  • Guidelines for RNA Purification from Whole Blood
  • Guidelines for RNA Purification from Yeast
  • Guidelines for Sample Stabilization Using the Monarch DNA/RNA Protection Reagent

• Application Notes

  • A Practical Guide to Analyzing Nucleic Acid Concentration and Purity with Microvolume Spectrophotometers
  • Facilitating Purification and Detection of Viral Nucleic Acids from Milk

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 https://www.neb.com/en/#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 P8107 in conjunction with the Monarch Total RNA Miniprep Kit?
  15. Can the Monarch Total RNA Miniprep Kit (NEB https://www.neb.com/en/#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 https://www.neb.com/en/#T2010) be used to purify RNA from plasma or serum?
  18. Can the Monarch Total RNA Miniprep Kit (NEB https://www.neb.com/en/#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 https://www.neb.com/en/#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 (https://www.neb.com/en/#M0303) with the Monarch Total RNA Miniprep Kit?
  25. I would like to purchase the contents of the Monarch Total RNA Miniprep Kit (NEB https://www.neb.com/en/#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 https://www.neb.com/en/#T2010), the Monarch gDNA Removal Columns (NEB https://www.neb.com/en/#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 https://www.neb.com/en/#T2017) and Monarch RNA Purification Columns (NEB https://www.neb.com/en/#T2007) included with the Monarch Total RNA Miniprep Kit (NEB https://www.neb.com/en/#T2010) compatible with the QIACube^® System?

• Troubleshooting

  • Troubleshooting Guide for Total RNA Extraction & Purification

Citations & Technical Literature

• Citations

  Product Citation Tool

  Additional Citations

  • 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 compatibility with QIACube has been updated.

  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]

  • T2010S_v1
  • T2010S_v2

• 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]

  • T2010S_v1_0011709
  • T2010G_v1_0031810
  • T2010S_v1_0031810
  • T2010G_v1_10029778
  • T2010S_v1_10029777
  • T2010G_v1_10043829
  • T2010S_v1_10043828
  • T2010G_v1_10044091
  • T2010S_v1_10044090
  • T2010G_v1_10053940
  • T2010S_v1_10053939
  • T2010S_v1_10068488
  • T2010G_v1_10068487
  • T2010S_v1_10075450
  • T2010S_v1_10079494
  • T2010S_v1_10081727
  • T2010S_v2_10095514
  • T2010G_v2_10097582
  • T2010S_v2_10097588
  • T2010G_v2_10141103
  • T2010S_v2_10141109
  • T2010G_v2_10144555
  • T2010S_v2_10144556
  • T2010G_v2_10144565
  • T2010S_v2_10144566
  • T2010G_v2_10166726
  • T2010S_v2_10166731
  • T2010G_v2_10175992
  • T2010S_v2_10175994
  • T2010S_v2_10214452
  • T2010S_v2_10214458
  • T2010S_v2_10214460
  • T2010G_v2_10210909
  • T2010S_v2_10210910
  • T2010S_v2_10247118

• Safety Data Sheets

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

  • Monarch^® DNA/RNA Protection Reagent
  • Monarch^® RNA Lysis Buffer
  • Monarch^® Proteinase K Reaction Buffer
  • Monarch^® DNase I Reaction Buffer
  • Monarch^® RNA Priming Buffer
  • Monarch^® RNA Wash Buffer
  • Monarch^® Nuclease-free Water
  • Monarch^® Proteinase K
  • Monarch^® Proteinase K Resuspension Buffer
  • Monarch^® DNase I

• 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.