Dynabeads™ mRNA Purification Kit (for mRNA purification from total RNA preps).

Citations & References (5)

Invitrogen™

Dynabeads™ mRNA Purification Kit (for mRNA purification from total RNA preps).

The Dynabeads mRNA Purification Kit is designed to specifically target, capture, and purify mRNA molecules from total RNA preparations.

Catalog Number	Quantity
61006	2 mL

Catalog number 61006

Price (BRL)

6.610,50

Add to cart

Quantity:

Price (BRL)

6.610,50

Add to cart

The Dynabeads™ mRNA Purification Kit rapidly isolates the mRNA transcriptome in typically 15 minutes, delivering pure, intact mRNA. The kit is designed to specifically target, capture, and purify mRNA molecules from total RNA preparations.

Advantages of using the Dynabeads™ mRNA Purification Kit:

Obtain pure mRNA in typically 15 minutes
Recover and enrich the transcriptome efficiently
Prepare mRNA that is suitable for nearly every downstream application

Pure, rapid mRNA isolation

The Dynabeads™ mRNA Purification Kit contains magnetic beads for the isolation of the mRNA transcriptome from any total eukaryotic RNA preparation (see figure). Dynabeads™ magnetic beads are uniform in size and highly mobile in solution (see figure). This enables the bead capture surface to quickly and continuously interact with the entire total RNA sample during the mRNA capture phase. The ribosomal RNA and small RNA molecules (transfer RNA, microRNA, small nucleolar RNA, and small cytoplasmic RNA) do not bind to the beads and are discarded. Only polyadenylated RNA species (mRNA) are captured resulting in cleaner, more sensitive results (see figure). Within about 15 minutes, pure mRNA is isolated and ready for use in downstream applications.

A straightforward enrichment procedure

The Dynabeads™ mRNA Purification Kit uses a robust affinity purification principle for the enrichment of polyadenylated mRNA. Superparamagnetic Dynabeads™, coupled to oligo-(dT)₂₅, are first equilibrated with Binding Buffer, and then mixed with purified total RNA. The beads are then washed to remove contaminating RNA species, and then mRNA is eluted in as little as 5 μL of 10 mM Tris-HCl. The entire process is facilitated by the use of a neodymium magnet (purchased separately), which allows for the quick and efficient immobilization of the magnetic beads during buffer changes. RNA is suitable for all downstream molecular applications, including:

Gene cloning
cDNA synthesis, cDNA library construction
RT-PCR, quantitative RT-PCR
RPA (Ribonuclease Protection Assay)
Subtractive hybridization
Dot/slot hybridization
Primer extension

For Research Use Only. Not for use in diagnostic procedures.

Specifications

Elution Volume5 to 20 μL

Final Product TypemRNA

For Use With (Application)RT-PCR, qPCR, cDNA library construction, microarray analysis

Green FeaturesBeads may be reused for multiple extractions

Purification Time10 min.

Quantity2 mL

Shipping ConditionRoom Temperature

Starting Material Amount≤100 μL

Yield2 μg mRNA per 200 μL of beads (Binding capacity)

Isolation TechnologyMagnetic Bead

Sample TypeTotal RNA

Unit SizeEach

Contents & Storage

• 2 mL Dynabeads Oligo (dT)₂₅; 4°C
• 5 mL Binding Buffer; 4°C
• 5 mL Washing Buffer B; 4°C
• 5 mL 10 mM Tris-HCl; 4°C

Frequently asked questions (FAQs)

I am getting DNA contamination after mRNA isolation using Dynabeads magnetic beads. Why is this?

There are several reasons why DNA contamination may occur:

- Incomplete DNA shearing.
- Incomplete removal of sample lysate after the hybridization step.
- Insufficient washing and/or removal of wash buffers.
- The ratio of sample to beads was too high.

My Dynabeads magnetic beads are not pelleting well with the magnet. Do you have any suggestions for me?

Please review the following possibilities for why your Dynabeads magnetic beads are not pelleting:

- The solution is too viscous.
- The beads have formed aggregates because of protein-protein interaction.

Try these suggestions: - Increase separation time (leave tub on magnet for 2-5 minutes)
- Add DNase I to the lysate (~0.01 mg/mL)
- Increase the Tween 20 concentration to ~0.05% of the binding and/or washing buffer.
- Add up to 20 mM beta-merecaptoethanol to the binding and/or wash buffers.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

I have a long double-stranded DNA fragment I would like to isolate. What product do you recommend?

For biotin-labeled DNA that is less than 1 kb, we recommend you use Dynabeads M270 Streptavidin (Cat. No. 65305) and MyOne C1 magnetic beads (Cat. No. 65001). We recommend our Dynabeads KilobaseBINDER Kit (Cat. No. 60101), which is designed to immobilize long (>1 kb) double-stranded DNA molecules. The KilobaseBINDER reagent consists of M-280 Streptavidin-coupled Dynabeads magnetic beads along with a patented immobilization activator in the binding solution to bind to long, biotinylated DNA molecules for isolation. Please see the following link (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/napamisc/capture-of-biotinylated-targets/immobilisation-of-long-biotinylated-dna-fragments.html) for more information in regards to long biotinylated DNA fragment isolation.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

Can I use Dynabeads magnetic beads to isolate single-stranded DNA templates?

Yes, Dynabeads magnetic beads can be used to isolate single-stranded DNA. Streptavidin Dynabeads magnetic beads can be used to target biotinylated DNA fragments, followed by denaturation of the double-stranded DNA and removal of the non-biotinylated strand. The streptavidin-coupled Dynabeads magnetic beads will not inhibit any enzymatic activity. This enables further handling and manipulation of the bead-bound DNA directly on the solid phase. Please see the following link (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/napamisc/capture-of-biotinylated-targets/preparing-single-stranded-dna-templates.html) for more information in regards to single-stranded DNA capture.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

What is the magnetic susceptibility for Dynabeads magnetic beads?

Magnetic susceptibility is a measure of how quickly the beads will migrate to the magnet. This will depend on the iron content and the character of the iron oxide. The magnetic susceptibility given for the Dynabeads magnetic beads is the mass susceptibility, given either as cgs units/g or m^3/kg (the latter being an SI unit). For ferri- and ferromagnetic substances, the magnetic mass susceptibility is dependent upon the magnetic field strength (H), as the magnetization of such substances is not a linear function of H but approaches a saturation value with increasing field. For that reason, the magnetic mass susceptibility of the Dynabeads magnetic beads is determined by a standardized procedure under fixed conditions. The magnetic mass susceptibility given in our catalog is thus the SI unit. Conversion from Gaussian (cgs, emu) units into SI units for magnetic mass susceptibility is achieved by multiplying the Gaussian factor (emu/g or cgs/g) by 4 pi x 10^-3. The resulting unit is also called the rationalized magnetic mass susceptibility, which should be distinguished from the (SI) dimensionless magnetic susceptibility unit. In general, magnetic mass susceptibility is a measure of the force (Fz) influencing an object positioned in a nonhomogenous magnetic field. The magnetic mass susceptibility of the Dynabeads magnetic beads is measured by weighing a sample, and then subjecting the sample to a magnetic field of known strength. The weight (F1) is then measured, and compared to the weight of the sample when the magnetic field is turned off (F0). The susceptibility is then calculated as K x 10^-3 = [(F1-F0) x m x 0.335 x 10^6], where K is the mass susceptibility of the sample of mass m. The susceptibility is then converted to SI units.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

View all Dynabeads™ mRNA Purification Kit (for mRNA purification from total RNA preps). FAQs

Citations & References (5)

Citations & References

Abstract

Identification of a novel glucose transporter-like protein-GLUT-12.

Authors:Rogers S, Macheda ML, Docherty SE, Carty MD, Henderson MA, Soeller WC, Gibbs EM, James DE, Best JD

Journal:Am J Physiol Endocrinol Metab

PubMed ID:11832379

'Facilitative glucose transporters exhibit variable hexose affinity and tissue-specific expression. These characteristics contribute to specialized metabolic properties of cells. Here we describe the characterization of a novel glucose transporter-like molecule, GLUT-12. GLUT-12 was identified in MCF-7 breast cancer cells by homology to the insulin-regulatable glucose transporter GLUT-4. The GLUT-12 cDNA ... More

Transcriptome analysis by strand-specific sequencing of complementary DNA.

Authors:Parkhomchuk D, Borodina T, Amstislavskiy V, Banaru M, Hallen L, Krobitsch S, Lehrach H, Soldatov A

Journal:Nucleic Acids Res

PubMed ID:19620212

High-throughput complementary DNA sequencing (RNA-Seq) is a powerful tool for whole-transcriptome analysis, supplying information about a transcript's expression level and structure. However, it is difficult to determine the polarity of transcripts, and therefore identify which strand is transcribed. Here, we present a simple cDNA sequencing protocol that preserves information about ... More

Transcriptome sequencing to detect gene fusions in cancer.

Authors:Maher CA, Kumar-Sinha C, Cao X, Kalyana-Sundaram S, Han B, Jing X, Sam L, Barrette T, Palanisamy N, Chinnaiyan AM

Journal:Nature

PubMed ID:19136943

Recurrent gene fusions, typically associated with haematological malignancies and rare bone and soft-tissue tumours, have recently been described in common solid tumours. Here we use an integrative analysis of high-throughput long- and short-read transcriptome sequencing of cancer cells to discover novel gene fusions. As a proof of concept, we successfully ... More

Widespread occurrence of antisense transcription in the human genome.

Authors:Yelin R, Dahary D, Sorek R, Levanon EY, Goldstein O, Shoshan A, Diber A, Biton S, Tamir Y, Khosravi R, Nemzer S, Pinner E, Walach S, Bernstein J, Savitsky K, Rotman G

Journal:Nat Biotechnol

PubMed ID:12640466

An increasing number of eukaryotic genes are being found to have naturally occurring antisense transcripts. Here we study the extent of antisense transcription in the human genome by analyzing the public databases of expressed sequences using a set of computational tools designed to identify sense-antisense transcriptional units on opposite DNA ... More

Phylogenomic analyses of lophophorates (brachiopods, phoronids and bryozoans) confirm the Lophotrochozoa concept.

Authors:Helmkampf M, Bruchhaus I, Hausdorf B

Journal:Proc Biol Sci

PubMed ID:18495619

Based on embryological and morphological evidence, Lophophorata was long considered to be the sister or paraphyletic stem group of Deuterostomia. By contrast, molecular data have consistently indicated that the three lophophorate lineages, Ectoprocta, Brachiopoda and Phoronida, are more closely related to trochozoans (annelids, molluscs and related groups) than to deuterostomes. ... More