Product Class: Kit

Monarch® Total RNA Miniprep Kit
Item# Description List Price Web Price Qty
T2010S Monarch Total RNA Miniprep Kit - 50 preps $333.00
$299.70
$233.10
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
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E6350L NEBNext rRNA Depletion Kit (Human/Mouse/Rat) with RNA Sample Purification Beads - 24 rxns $1,775.00
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E6560L ProtoScript II First Strand cDNA Synthesis Kit - 150 rxns $921.00
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E6560S ProtoScript II First Strand cDNA Synthesis Kit - 30 rxns $229.00
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E7760S NEBNext Ultra II Directional RNA Library Prep Kit for Illumina - 24 rxns $1,356.00
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E7765L NEBNext Ultra II Directional RNA Library Prep with Sample Purification Beads - 96 rxns $5,111.00
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E7765S NEBNext Ultra II Directional RNA Library Prep with Sample Purification Beads - 24 rxns $1,501.00
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E7770L NEBNext Ultra II RNA Library Prep Kit for Illumina - 96 rxns $4,386.00
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M0368L ProtoScript II Reverse Transcriptase - 10,000 units $246.00
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    Product Introduction

    Quickly and easily purify up to 100 µg of high quality total RNA from multiple sample types – all with one kit!

    • For use with blood, cells and tissues
    • 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)
    • Contains Proteinase K for processing of tissues and blood samples
    • Includes RNA Protection Reagent for sample preservation
    • Excellent value
    • Kit components available separately

    Product Information

    Description

    request monarch sample

    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

    Specifications:

    Binding Capacity: 100 µg RNA
    RNA Size: ≥ 20 nt
    Purity: A260/280 usually ≥ 1.8, 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 107
    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

    Validated 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.

     

    Kit Components

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    Monarch® Proteinase K 25
    Monarch® Proteinase K Resuspension Buffer 25
    Monarch® Proteinase K Reaction Buffer 25
    Monarch® DNase I 25
    Monarch® DNase I Reaction Buffer 25
    Monarch® Nuclease-free Water 25
    Monarch® RNA Purification Columns 25
    Monarch® DNA/RNA Protection Reagent 25
    Monarch® RNA Lysis Buffer 25
    Monarch® RNA Priming Buffer 25
    Monarch® RNA Wash Buffer 25
    Monarch® gDNA Removal Columns 25
    Monarch® Collection Tubes II 25
    Product Categories:
    RNA Reagents Products,
    Total RNA Extraction & Purification Products,
    RNA Extraction & Purification Products
    ,
    Nucleic Acid Purification Products
    Applications:
    RNA Analysis,
    RNA Purification and Isolation

    Properties & Usage

    Storage Temperature

    25°C

    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 Monarch® Total RNA Miniprep Kit (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. Total RNA Purification from Samples in RNAlater® 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 Stored in Monarch DNA/RNA Protection Reagent using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    9. FFPE Tissue Deparaffinization and Subsequent RNA Purification Using the Monarch Total RNA Miniprep Kit NEB T2010
    10. In-tube DNase I Treatment after Purification with the Monarch Total RNA Miniprep Kit (NEB #T2010)
    11. RNA Reaction Cleanup using the Monarch Total RNA Miniprep Kit (NEB #T2010)
    12. Fractionation of Small and Large RNA Using the Monarch Total RNA Miniprep Kit (NEB #T2010)

    Manuals

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

    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

    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. How can I tell the difference between the gDNA Removal Column and the RNA Purification Column?
    5. Can the Monarch® Total RNA Miniprep Kit be used to purify RNA from yeast?
    6. Can the Monarch® Total RNA Miniprep Kit be used to purify RNA from bacteria?
    7. Can the Monarch® Total RNA Miniprep Kit be used to purify RNA from plants?
    8. How can I preserve my sample if I am not ready to extract RNA right after harvest?
    9. Can I purchase Monarch® buffers and columns separately?
    10. I am seeing genomic DNA in my preps when using the Monarch Total RNA Miniprep Kit. What could be the problem?
    11. Can I use the Monarch® Total RNA Miniprep Kit for RNA cleanup after enzymatic manipulations?
    12. Can I skip the on-column DNase I treatment step of the Monarch® Total RNA Miniprep Kit protocol?
    13. Can I skip the gDNA removal column step of the Monarch® Total RNA Miniprep Kit protocol?
    14. Can I use leftover Proteinase K and DNase I from the Monarch® Total RNA Miniprep Kit for other applications?
    15. Can I use NEB Proteinase K, Molecular Biology Grade (NEB #P8107S) in conjunction with the Monarch® Total RNA Miniprep Kit?
    16. Can I use NEB DNase I (#M0303) with the Monarch Total RNA Miniprep Kit?
    17. Can the Monarch Total RNA Miniprep Kit (NEB #T2010) be used to purify RNA from saliva?
    18. Can the Monarch Total RNA Miniprep Kit NEB T2010 be used to purify RNA from buccal (cheek) swabs?
    19. Can the Monarch Total RNA Miniprep Kit (NEB #T2010) be used to purify RNA from plasma or serum?
    20. Can the Monarch Total RNA Miniprep Kit (NEB #T2010) be used to purify RNA from nucleated blood (e.g. bird, fish, reptile)?
    21. Can the Monarch Total RNA Miniprep Kit be used to purify RNA from drosophila?
    22. Can the Monarch Total RNA Miniprep Kit be used to purify RNA from zebrafish embryos/larvae?
    23. Can the genomic DNA captured on the Monarch gDNA Removal Column (NEB #T2017) be eluted and used in downstream applications?
    24. Can I lyse my adherent cells directly in their wells when using the Monarch Total RNA Miniprep Kit?
    25. How can I assess RNA integrity and purity?

    Troubleshooting

    • Troubleshooting Guide for Total RNA Extraction & Purification

    Quality & Safety

    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

    This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

    While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

    For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

    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.