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Cat. No.: O25MB-1 (for 1ml)

Cat. No.: O25MB-5 (for 5ml)

Cat. No.: O25MB-20 (for 20ml)

Cat. No.: O25MB-100 (for 100ml)

Description

Oligo (dT)25 Magnetic Beads (mRNA Magnetic Beads), also known as mRNA Magnetic Beads or Oligo dT Magnetic Beads, are magnetic beads coated with Oligo (dT)25. These beads can complementarily pair with the poly(A) tails of mRNA, allowing for the stable, efficient, and convenient direct separation and purification of high-purity intact mRNA and other RNA with poly(A) tails from animal tissues, cultured animal cells, or total RNA.

The purified mRNA and lncRNA with poly(A) tails obtained using this product can be directly used in RT-PCR, qPCR, high-throughput sequencing, mRNA library construction, m6A modification analysis, solid-phase cDNA library construction, Northern blot analysis, RACE, and other molecular biology experiments. It can also be used in mRNA vaccine research, such as for the purification of mRNA vaccines or the separation and purification of short poly(A) tails during 3' Poly A tail length detection.

The poly(A) RNA purified by this product may also include lncRNA with poly(A) tails, but it is primarily mRNA, which is why it is often referred to as mRNA Magnetic Beads.

In a typical mammalian cell, the four main macromolecules and their approximate mass and percentage are: RNA, ~20 pg (1%); DNA, ~7 pg (0.3%); protein, ~500 pg (20%); polysaccharide, ~2 μg (78.7%). Messenger RNA (mRNA) accounts for approximately 4% of the total RNA mass, while ribosomal RNA (rRNA) accounts for about 80%.

Principle and main operation process: The surface of Oligo (dT)25 Magnetic Beads is covalently modified with 25-mer dT sequences, i.e., Oligo (dT)25. When lysates from eukaryotic cells, plant or animal tissues, or extracted total RNA are mixed with Oligo (dT)25 Magnetic Beads, the oligo dT sequences on the bead surface base-pair with the 3' poly(A) tails of mRNA and other RNAs for specific binding. Under the influence of an external magnetic field, the beads can quickly and efficiently separate from the corresponding solution. After thorough washing to remove impurities, the mRNA and other poly(A) RNAs are eluted from the beads using an elution buffer, resulting in high-purity intact mRNA and other poly(A) RNAs.

Features

• This product offers stable extraction performance, high purity, fast processing speed, and convenient operation. The mRNA extraction system of this product has been repeatedly tested and optimized, capable of purifying over 90% of mRNA from total RNA. It can extract approximately 0.1-1 µg of mRNA from 10^6 cells, about 0.2-2 µg of mRNA from 20 mg of mouse liver tissue, and around 0.1-1 µg of mRNA from 20 mg of mouse lung tissue. The entire purification process, which involves simple steps of lysis, binding, washing, and elution, can be completed within 15 minutes. All operations are carried out in a single centrifuge tube, making it highly convenient.
• The magnetic beads in this product are approximately 200 nm in diameter, with a concentration of about 5 mg/ml. Each milligram of beads is conjugated with approximately 300-400 pmol of Oligo (dT)25. Each milligram of beads can purify approximately 2-3 µg of mRNA or other poly(A) RNA, which means that each milliliter of beads can purify about 10-15 µg of mRNA or other poly(A) RNA.

Precautions

• Ensure RNase-free and DNase-free conditions during the operation. Use RNase-free and DNase-free reagents and consumables, and handle them carefully to avoid contamination. If consumables might be contaminated with RNase, soak them overnight in 0.01% DEPC water, then autoclave and dry them. High-temperature autoclaving can deactivate DNase if contamination is possible.
• A magnetic separation device is required, which we can also provide.
• When dispensing or using the beads, gently vortex or invert the tube several times to ensure the beads are well mixed.
• Before magnetic separation, gently shake the centrifuge tube to disperse the beads fully, then bring it close to the magnetic field. If the beads adhere to the tube wall, shake the liquid in the tube after bead aggregation to let the adhering beads fall off.
• Use the recommended sample volume. Excessive sample volume may cause bead aggregation, affecting washing and the purity of the extracted mRNA. If bead aggregation occurs, disperse the beads as much as possible during washing to improve extraction efficiency. If bead aggregation persists, reduce the total sample volume in subsequent experiments.
• Since mRNA degrades easily, it is recommended to use the extracted mRNA promptly for RT-PCR or other experiments. If immediate use is not possible, store it at -80ºC.
• This product is for scientific research use by trained professionals only. It is not for clinical diagnosis or treatment, food, or drug use, and should not be stored in residential areas.
• For your safety and health, wear a lab coat and disposable gloves while handling.

Storage

Store at 4ºC, valid for one year. For long-term storage, Oligo (dT)25 Magnetic Beads can be stored at -20ºC, which will extend their shelf life further.

Only for research and not intended for treatment of humans or animals

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SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory