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RNA Purification and Isolation

Isolating high-quality RNA molecules is crucial to many downstream experiments, such as cloning, reverse transcription for cDNA synthesis, RT-PCR, RT-qPCR and RNA-seq. There are various approaches to RNA purification including phenol-chloroform extraction, spin column purification, and the use of magnetic beads.

Total RNA purification involves the extraction and purification of total RNA from your sample, for use in gene expression analyses such as RT-qPCR or RNA-seq. Depending on the starting sample, different protocols should be employed to most efficiently lyse cells or tissues, and purify the RNA from contamination genomic DNA and other cellular components. Theoretically, total RNA purification involves the isolation of all of the cellular RNA, including various sizes ranging from intact ribosomal RNA down to small RNA (including miRNA, tRNA and 5sRNA).

For recovery of a specific RNA subpopulation from total RNA derived from cells, tissues or in vitro transcriptions, bead-based methods may be utilized to isolate mRNAs through binding of the poly(A) tail or to isolate RNAs based on size by varying the ratio of nucleic acid to beads in the presence of a crowding reagent. Bead-based methods are a rapid way to isolate RNA free from enzymes, salts and nucleotides from a variety of sources, and when RNA is present at lower concentrations.

In vitro transcribed RNA also often needs to be cleaned up and separated from components such as DNA templates, unincorporated nucleotides or RNA modifying enzymes. Chemical-, column- and gel-based approaches can be used, and are sometimes combined to result in purified RNA free from nucleotides, salts and proteins. Learn more about cleaning up in vitro transcribed RNA. (hyperlink “Learn more” to child page “Cleaning up in vitro transcribed RNA”)

Regardless of the methods employed, purified RNA is ready for use in various downstream applications including expression analysis, transient transfections or microinjections to assess genome editing or RNA localization, as well as in vitro assays including RNA structure studies, nucleic acid hybridizations and in vitro translation.

 

FAQs for RNA Purification and Isolation

Protocols for RNA Purification and Isolation

    Other Tools & Resources

    Brochures

  • Monarch Nucleic Acid Purification Brochure

    It's time to transform your DNA purification experience! Learn about the advantages of choosing Monarch Nucleic Acid Purification Kits from NEB.

Reagents for RNA Sample Preparation

NEB offers a wide variety of reagents to facilitate front-end processing of RNA for downstream applications including next-generation sequencing, library construction and microarrays.

RNA Polymerase Selection Chart

NEB offers RNA polymerases that can be used for in vitro synthesis of RNA for a wide variety of downstream applications. Additional polymerases are offered for the generation of untemplated homoribopolymeric tails for reverse transcription or labeling.

RNA Ligase Selection Chart

NEB offers a variety of ligases for RNA research with unique specificities.

cDNA Synthesis Selection Chart

NEB offers several reagents for cDNA synthesis for use in applications including qPCR and qRT-PCR. For your convenience, reagents are available as kits or standalone products, depending on your needs.

Legal Information

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 gbd@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.

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    How to recycle your Monarch Kit components

    Learn how you can easily recycle all of the components in your Monarch Kits.