RNA Extraction and Purification

Isolation of RNA molecules is crucial to many molecular biology applications, such as cloning, reverse transcription for cDNA synthesis, RT-PCR, RT-qPCR and RNA sequencing (RNA-seq). There are various approaches to RNA extraction and purification including phenol-chloroform extraction, spin column purification, and magnetic bead-based methods. Methods are selected based on sample characteristics, contaminant types, target RNA length and abundance, scale of sample processing and downstream analyses.

The Monarch Total RNA Extraction and RNA Cleanup Kits feature silica spin column-based RNA isolation protocols to achieve desired RNA yields, purity, and integrity with a wide variety of sample types. The Monarch RNA workflows have multiple capacity options and are also amendable to automated RNA extraction. Monarch kits and individual components are only available in sustainably designed formats that avoid excess waste associated with conventional RNA isolation kits and protocols.

Total RNA purification involves the extraction and purification of total RNA from your sample. Depending on the starting sample, different protocols should be employed to lyse cells or tissues and purify the RNA from contaminant genomic DNA and other cellular components most efficiently. Theoretically, total RNA purification involves the isolation of all the cellular RNA, including various sizes ranging from intact ribosomal RNA down to small RNA (including miRNA, tRNA and 5sRNA). Silica column-based RNA extraction methods are adaptable for fractionation of small and large RNA species.

The Monarch RNA Total Miniprep Kit is a universal kit for RNA extraction and purification from a wide variety of cell types and tissues, including blood, saliva, FFPE, cultured cells, viral, and tough-to-lyse samples (including plant, bacterial and yeast). The kit includes a protection reagent for sample storage and lysis, Proteinase K for homogenization of some sample types, gDNA removal columns and DNase I treatment after extraction for genomic DNA removal.

Enzymatic reactions in which RNA is used as a substrate may require cleanup steps for buffer exchange and removal of contaminants. In vitro transcribed (IVT) RNA often needs to be cleaned up and separated from components that are considered contaminants in downstream applications, 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.

The Monarch RNA Cleanup Kits provide a fast and simple silica column-based solution for cleanup and concentration of RNA after enzymatic reactions (including purification of in vitro transcription (IVT), DNase I and Proteinase K treatment, capping, tailing, and labeling) as well as after RNA isolation (e.g., TRIzol^® RNA extraction). 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.