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RNA Purification and Detection

Isolating high-quality RNA is crucial to many downstream application, such as cloning, reverse transcription for cDNA synthesis, RT-PCR, RT-qPCR and RNA-seq. Isolation of total RNA from cells, blood, tissues and other samples can be accomplished using guanidium-phenol reagents followed by precipitation with lithium chloride and ethanol. Alternatively, commercial kits containing optimized buffer systems and silica columns can be employed.
 
Studies that require the isolation of eukaryotic mRNA often take advantage of the polyadenine tail typically present at the 3’ end. One approach involves the use affinity matrices; poly-T oligonucleotides that hybridize to the complementary poly-A tails (1). Enrichment of miRNA and siRNAs can also be achieved using high-affinity binding proteins and the use of chitin magnetic beads (2, 3).
 
Similar to DNA, enzymatic reactions in which RNA is used as a substrate may require cleanup steps for buffer exchange or removal of contaminants. Commercial RNA cleanup kits are also available for this application.

(1) Aviv, H. and Leder, P. (1972) Proc. Natl. Acad. Sci. USA, 69, 1408. PMID: 4504350 
(2) Silhavy, D. et al. (2002) EMBO J., 21, 3070-3080. PMID: 12065420 
(3) Jin, J. et al. (2010) Biotechneques. 48, XVII-XXII. PMID: 20569217

FAQs for RNA Purification and Detection

Protocols for RNA Purification and Detection

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.