96 mRNA Isolation Kit

Supplier
Ademtech
Cat. No
2006031
Kit size
96 preps
Links
ademtech Protocol Information Sheet

Features

• Complete kit adapted for Total RNA, Cells or whole blood
• Microplate format: convenient for many mRNA isolation
• More convenient for many subsequent RT-PCR
• Very low ribosomal contamination
• Intact and pure mRNA without degradation
• Purification of all mRNA sizes
• Fast system: less than 30 minutes

mRNA purification procedure relies on the base pairing between the poly A tail of the mRNA and the biotinylated oligonucleotides (dT)25 sequence bound to the Bio-Adembeads Streptavidin. Nucleo-Adembeads consists of superparamagnetic nanoparticles of uniform size and a perfect spherical shape. Oligonucleotides (dT)25 are linked to the surface of the superparamagnetic nanoparticles via Biotin-Streptavidin system. The true spherical shape eliminates n-specific binding associates with irregulary shape particles. After processing the protocol using Nucleo-Adembeads, the mRNA is ready to use for downstream applications such as RT-PCR, Nothern Blotting, cDNA library construction, nuclease protection assay, in vitro translation, primer extension, subtractive cDNA cloning and reverse transcription.

• Prepare samples adequately with protocol (Total RNA, Cell lysate or whole blood)
• Add prepared samples solutions to the [Oligo (dT)25 - Bio-Adembeads Streptavidin] complex and mix by pipetting.
• Incubate for 10 minutes at room temperature.
• Place the plate on the magnet, remove and discard the supernatants. Move off the plate and wash with: - 100 μl Washing Buffer for mRNA purification starting from Total RNA. Repeat twice this step. - 150 μl Lysis Buffer for mRNA purification starting from cells or whole blood.Repeat twice this step using 200μl of Washing Buffer.
• Place the plate on the magnet (also available from Labomics), remove and discard the supernatants.
• Move off the plate and resuspend the complexes in desired volume of Nuclease free Water (mix by pipetting).
• After 2 minutes at room temperature, place the plate on the magnet during 5 minutes and transfer the supernatants (containing the mRNA) into a collecting plate for storage.

Beads	1 mL Bio-Adembeads Streptavidin
Buffers	30 mL Lysis Buffer	15 mL Binding Buffer 3X	40 mL Washing Buffer
Other	15 mL Nuclease free Water	Lyophilisate Biotinylated Oligo(dT)25
Storage	Store at 2-8°C.

• mRNA purification from up to 25 µg Total RNA, 10⁵ cells from Cell lysate or 25 µL whole blood