This mRNA isolation kit specifically targets and captures mRNA molecules from virtually any crude sample and eliminates the need to purify total RNA when the desired information bearing nucleic acid is mRNA. Since the mRNA content of cellular total RNA is only ~1-5%, why isolate total RNA when just a sub-fraction of the total RNA component is mRNA? Other 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 membrane binding.
- 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 DIRECT™ Kit specifically target and capture the mRNA transcriptome from an extremely wide variety of crude starting samples. Ribosomal RNA, DNA, proteins, and small RNA molecules (such as transfer RNA, micro RNA and small nucleolar 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 or a post-extraction DNase treatment.
RNase inhibitior agents in the Lysis/Binding buffer together with stringent hybridization and washing steps ensure that the mRNA isolation yields intact mRNA even from samples that are rich in RNA degrading enzymes. Careful buffer formulations prevent RNA degradation without the use of strong chaotropic agents.
Designed for an incredibly broad range of sample types:
- 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
- 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
Verastile Elution Options:
- Elution in any volume down to 5 µl
- 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