Product Class: Kit

Monarch® Spin RNA Isolation Kit (Mini)


Catalog #T2110

Product Introduction

  • A new version of total RNA extraction ‘miniprep’ kit, featuring upgraded spin columns precision-engineered for high performance
  • Purify up to 100 µg of high-quality total RNA from multiple sample types including tough-to-lyse samples with this single all-in-one kit
  • Extract RNA from samples such as cells, tissues including fibrous or lipid-rich, Gram-positive or Gram-negative bacteria, yeast, plants, insects and more; kit includes Proteinase K for sample processing and Monarch StabiLyse™ DNA/RNA Buffer for sample preservation
  • Achieve binding capacity of a standard mini kit (100 µg) with the low elution volume of a micro kit (10 µl)
  • Remove genomic DNA contamination with specialized columns and RNase-free DNase included in the kit
  • Capture total RNA pool in your sample including small RNAs <200 nt

Product Box for T2110

Product Information

Description

The Monarch Spin RNA Isolation Kit (Mini) is a comprehensive solution for sample preservation, cell lysis, gDNA removal, and purification of total RNA from various biological samples, including cultured cells, mammalian tissues, microbes, plants, insects and blood. With just this single kit, users can purify up to 100 µg of high-quality total RNA from multiple sample types, from standard as well as tough-to-lyse samples, with included enzymes, reagents, and specialized gDNA removal columns. The kit uniquely enables binding capacities like RNA purification miniprep kits, combined with the low elution volumes of micro kits. Cleanup of enzymatic reactions and purification of RNA from TRIzol® -extracted samples is also possible using this kit. Purified RNA has metrics with A260/280 and A260/230 ratios typically > 2.0, high RNA integrity scores, and minimal residual gDNA. This kit captures RNA ranging in size from full-length rRNAs down to intact miRNAs. Purified RNA is suitable for downstream applications such as RT-qPCR, cDNA synthesis, RNA-seq, and RNA hybridization-based technologies.

Designed with sustainability in mind, Monarch kits use significantly less plastic than other kits on the market.

Properties

Purification Format

Spin column for nucleic acid purification

Compatible Methods

Centrifugation or vacuum manifold

Intended Usage

Total RNA isolation

Compatible Sample Types

Cells, microbes, plants, insects, mammalian tissues including soft, fibrous and lipid-rich, blood, plasma

Yield

Varies depending on sample type. Refer to Choosing Sample Input Types for RNA Purification and Average Yield Expected.

Column Binding Capacity

Up to 100 μg total RNA

RNA Size

≥ 20 nt, total RNA including small RNA, miRNA, mRNA, lncRNA, rRNA

Elution Volume

≥ 10 μl

RNA Purity

Typically A260/280 ≥ 2.0  and A260/230 ≥ 1.8

Compatible
Downstream Applications

cDNA synthesis, RT-qPCR, RNA-Seq, small RNA library prep, Northern blotting, hybridization-based workflows

 

 

Figure 1: Overview of the Monarch Spin RNA Isolation Kit workflow

Workflow for the Monarch Spin RNA Isolation Kit

The Monarch Spin RNA Isolation Kit (Mini) enables total RNA purification from a wide range of sample types and elution in low volumes. The kit features unique spin columns designed with less plastic and includes reagents for optimal RNA extraction.


Figure 2: Monarch Spin RNA Isolation Kit (Mini) is a versatile solution for high-quality RNA extraction from a wide variety of sample types

Gel showing a variety of samples



RNA was extracted in duplicate using the Monarch Spin RNA Isolation Kit (Mini) from various samples representing a range of biological properties in the starting material, including fibrous, fatty and nuclease-rich mammalian tissues, tough-to-lyse samples such as Gram-positive bacteria, plants and insects, as well as RNA cleanup reactions. RNA quality was assessed using A260/A280 and A260/A230 ratios measured using a microvolume spectrophotometer (Lunatic®, Unchained Labs®). RNA integrity was measured using Agilent® automated electrophoresis systems according to sample types. For samples with known typical RNA profiles, Agilent RNA ScreenTape® was used on the TapeStation® 4200 (A). For samples with known atypical RNA profiles such as plant green tissue with plastidal content, and insects with ribosomal gene breaks, Agilent Bioanalyzer® 2100 used with NanoChip (B).


Figure 3: Monarch Spin RNA Isolation Kit (Mini) enables extraction of high-quality RNA even from low input samples

Gel and qPCR data for a variety of inputs (10^2 to 10^6 cells)

RNA was extracted using the Monarch Spin RNA Isolation Kit (Mini) from varying amounts of HEK293 cells over 5 orders of magnitude starting from 1 million cells to 100.
A. To assess RNA integrity, the extracted RNA was resolved using the Agilent® Pico Bioanalyzer chip on a Bioanalyzer 2100 with sample loading adjusted to the manufacturer's recommendation of assay input range.
B. To demonstrate successful downstream application, the RNA was subjected to RT-qPCR using Luna® Probe One-Step RT-qPCR 4X Mix with UDG (NEB #M3019) in a 20 μl reaction targeting GAPDH mRNA. Expected amplification curves were observed showing a quantitative trendcorresponding to the varying input amounts.


Figure 4: Monarch Spin RNA Isolation Kit (Mini) successfully purifies small RNAs below 200 nucleotides, enabling a more faithful representation of the total RNA pool compared to a leading supplier

Data showing comparison to Supplier Q for Small RNA

RNA was extracted from 10 mg mouse brain tissues using Monarch Spin RNA Isolation Kit (Mini) and a comparable kit from Supplier Q. To visualize the isolation of small RNA species, the extracted RNA was resolved using the Agilent Small RNA chip on a Bioanalyzer 2100 according to manufacturer's instructions. Gel traces of RNA isolated from the two kits are shown (A), with representative electropherograms showing the small RNA size profiles (B).


Figure 5: Monarch Spin RNA Isolation Kit (Mini) offers greater flexibility in elution volumes with elution as low as 10 μl

Bar graph showing RNA yield versus elution volume

RNA was extracted in replicate using the Monarch® Spin RNA Isolation Kit (Mini) with an input of 1 million HEK293 cells per extraction. To assess the recovery of RNA at varying elution volumes, RNA was eluted in 100, 50 30, 20 or 10 μl nuclease-free water. RNA concentration and subsequent yield were measured using a microvolume UV-Vis spectrophotometer (Lunatic®, Unchained Labs®). An elution volume of at least 20 μl is recommended to achieve maximal RNA recovery however, elution in as low as 10 μl can be applied depending on user's needs.


 

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

Protocols, Manuals & Usage

Protocols

  1. Protocol for Total RNA Isolation and Purification Using the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110)
  2. Protocol for Sample Lysis of Bacteria or Yeast Using the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110)
  3. Protocol for Sample Lysis of Cells Using the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110)
  4. Protocol for Sample Lysis of Plants or Insects Using the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110)
  5. Protocol for Sample Lysis of Tissues or Leukocytes Using the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110)
  6. Protocol for RNA Reaction Cleanup Using the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110)

Manuals

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

Usage & Guidelines

FAQs & Troubleshooting

FAQs

  1. Is the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110) compatible with Luna RT-qPCR Reagents?
  2. Is the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110) compatible with NEBNext reagents for RNA Library Prep?
  3. What is the lowest volume in which I can elute my RNA with the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110)?
  4. When using the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110), how can I preserve my sample if I am not ready to extract RNA right after harvest?
  5. When reconstituting Buffer BX from the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110), does it matter if I add water or ethanol first?
  6. When using Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110), upon ethanol addition to the flow-through, the sample looks cloudy. Is this a problem?
  7. When using Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110), the addition of ethanol to the flow-through results in a white insoluble precipitate. Should I load this onto the column or leave it out?
  8. I see genomic DNA in my preps when using the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110). What could be the problem?
  9. Can I use the Monarch Spin RNA Isolation Kit (Mini) for RNA (NEB #T2110) cleanup after enzymatic manipulations?
  10. Can I skip the on-column DNase I treatment step of the Monarch Spin RNA Isolation Kit (Mini) protocol (NEB #T2110)?
  11. Can I skip the gDNA removal step with the gDNA removal column (Monarch Spin Column S2C) of the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110) protocol?
  12. Can I use leftover Proteinase K and DNase I from the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110) for other applications?
  13. Can I use NEBExpress T4 Lysozyme (NEB #P8115) with the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110)?
  14. Can the genomic DNA captured on the Monarch Spin Columns S2C (NEB #T3017) be eluted and used in downstream applications?
  15. Can I lyse my adherent cells directly in their wells when using the Monarch Spin RNA Isolation Kit (Mini) (NEB #T2110)?
  16. How can I assess RNA integrity and purity?
  17. Can I use NEB DNase I (NEB #M0303) with the Monarch Spin RNA Isolation Kit?

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.

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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