Dynabeads® mRNA Purification Kit for mRNA Purification from Total RNA preps

Isolate the Complete mRNA Transcriptome in 15 minutes. This mRNA isolation kit specifically targets, captures and purifies mRNA molecules from total RNA preparations. Since the mRNA content of eukaryotic total RNA preparations is only ~1-5% we recommended isolating mRNA directly from crude lysates when possible (see: Dynabeads® Direct mRNA Isolation kits). This is because technologies designed to purify total RNA yield ~80% ribosomal RNA and force mRNA to compete with ribosomal RNA, transfer RNA, micro RNA, small nucleolar RNA and small cytoplasmic RNA for purification.

• 15 minute procedure yields pure intact mRNA
• Extremely pure mRNA isolation, best choice upstream of cDNA synthesis
• Exquisitely sensitive mRNA isolation enables cDNA synthesis and cDNA library construction from ultra-small starting samples. (Enables cDNA library construction from a single cell).

The mRNA purification beads in the Dynabeads® mRNA Purification Kit specifically target, capture and purify mRNA transcriptome from preparations of total eukaryotic RNA from any source. Ribosomal RNA and small RNA molecules (such as transfer RNA, micro RNA and small nucleolar RNA and small cytoplasmic RNA) do not bind to the beads and are discarded. Only polyadenylated RNA species (mRNA) are captured. Isolated mRNA is pure, eliminating the need for ribosomal RNA subtraction.

Purify mRNA from total RNA preps of any eukaryotic origin:

• Total RNA purification prep
Blood, serum, plasma, sputum
• Viral poly A+ RNA from cells, serum, plasma
• Mammalian, amphibian, fish, and insect tissues: Brain, Lung, Liver, Gut, Heart, Kidney, Pituitary, Pancreas, Ovary, Muscle, Hypothalamus, Eggs, Whole Insect, etc...
• Plant tissues: Whole Plants, bud, guard cell (single cells), embryos, flowers, leaves, ovules, roots, seed (aleurone, endosperm), stigma, etc...
• Formalin-fixed, paraffin-embedded tissues
• Yeast (Sacchromyces cerevisiae)
• Unfractionated nucleic acid sample (DNA + Total RNA)

mRNA is suitable for all downstream molecular applications:

• gene cloning
• cDNA synthesis, cDNA library construction
• RT-PCR, Quantitative RT-PCR
• RPA - Ribonuclease Protection Assay
• Subtractive Hybridization
• Dot/slot Hybridization
• Primer extension
• SAGE, RACE, etc...

Column-free system ensures highest transcriptome recovery:

• Physical mRNA capture on mobile magnetic beads
• Rapid and gentle magnetic handling procedures
• No mRNA lost during high g-force spins
• No mRNA trapped in column membranes during elution

The ideal mRNA purification method prior to cDNA library construction

• Ensures the highest recovery and enrichment of the transcriptome
• Capture more of the transcriptome than is possible with methods that integrate a total RNA isolation step upstream of mRNA isolation

Dynabeads are highly mobile in solution enabling the mRNA capture surface to freely and continuously interact with the entire sample volume during the mRNA capture phase. Strong magnetic fields quickly pull the Dynabeads to the tube wall, allowing for easy and complete removal of the supernatant by pipette. Washing steps are performed similarly. Pure mRNA is eluted free of other contaminating RNA species.

Verastile Elution Options:

• Elution in any volume down to 5µl
• mRNA elution is optional:
    – Enzymatic reactions in downstream procedures are not inhibited by presence of Dynabeads
    – Can perform cDNA synthesis directly on the beads to create a reusable solid-phase cDNA library