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Cat. No.: AMPKMB-50 (for 50T)

Cat. No.: AMPKMB-200 (for 200T)

Cat. No.: AMPKMB-800 (for 800T)

Description

Animal mRNA Isolation Kit with Magnetic Beads is a reagent kit that utilizes Oligo (dT)25-coated magnetic beads and an optimized buffer system for stable, efficient, and convenient isolation and purification of high-purity, intact mRNA from animal tissues or cultured animal cells.

The mRNA extracted using this kit can be directly used in molecular biology experiments such as RT-PCR, qPCR, high-throughput sequencing, mRNA library construction, solid-phase cDNA library construction, Northern blot analysis, RACE, and other applications, including mRNA vaccine development.

Although the poly(A) RNA extracted by this product may also contain poly(A)-containing lncRNAs and other transcripts, it predominantly consists of mRNA, hence referred to as an mRNA extraction kit.

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

The Oligo (dT)25 magnetic beads are covalently modified with a 25-mer poly(dT) sequence, which specifically binds to the poly(A) tail at the 3' end of mRNA through base pairing when mixed with the lysate of eukaryotic cells or animal/plant tissues. Subsequently, under the influence of an external magnetic field, the magnetic beads can be rapidly and efficiently separated from the corresponding solution, thoroughly washed to remove impurities, and finally, the mRNA is eluted from the beads using an elution buffer, resulting in the obtainment of high-purity, intact mRNA.

Components

50T

• Oligo (dT)25 Magnetic Beads    1ml
• Solution I (Binding Buffer)    30ml
• Solution II (Lysis Buffer)    30ml
• Solution III (Wash Buffer I)    30ml
• Solution IV (Wash Buffer II)    60ml
• Solution V (Elution Buffer)    5ml

Advantages

The kit offers several advantages, including stable extraction performance, high purity, fast speed, and convenient operation. The mRNA extraction system of this kit has been extensively tested and optimized. It can extract approximately 0.1-1 µg of mRNA from 106 cells, approximately 0.2-2 µg of mRNA from 20 mg of mouse liver tissue, and approximately 0.1-1 µg of mRNA from 20 mg of mouse lung tissue. The entire purification process can be completed in less than 15 minutes, requiring only simple steps such as incubation, washing, and elution. All the operations can be carried out in a single centrifuge tube, ensuring convenience.

Product Information

Oligo (dT)25 magnetic beads in this kit have a particle size of approximately 200nm and a concentration of approximately 5mg/ml. Each milligram of magnetic beads is coupled with approximately 300-400 pmol of Oligo (dT)25, and can purify approximately 2-3 µg of mRNA. Each milliliter of magnetic beads can purify approximately 10-15 µg of mRNA.

For routine mRNA extraction, it is recommended to use 20 μl of magnetic bead suspension per sample. The small packaging of this kit can be used for 50 mRNA extractions, the medium packaging can be used for 200 mRNA extractions, and the large packaging can be used for 800 mRNA extractions.

Precautions

• Please ensure strict measures to prevent contamination with RNAases and DNAases during the operation.
• All reagents and consumables used in this product must be RNase-free and DNase-free. Exercise caution during the procedure to avoid contamination. If there is a possibility of RNase contamination in the consumables, consider soaking them overnight in 0.01% DEPC-treated water, followed by high-temperature and high-pressure sterilization, and drying. High-temperature and high-pressure sterilization is generally effective in inactivating DNases if there is a possibility of DNase contamination.
• A magnetic separation device is required for use and should be obtained separately. We recommend using our VSep™ Magnetic Separators.
• When aliquoting or working with the magnetic beads, it is recommended to vortex or invert them gently to ensure thorough mixing.
• Before magnetic separation, gently agitate the centrifuge tube to ensure even dispersion of the magnetic beads before bringing them close to the magnetic field. If the magnetic beads stick to the tube wall, gently shake the liquid in the tube to make the beads flow down after they aggregate.
• Please use the recommended amount of total RNA. If the total RNA amount is too high, it may cause bead aggregation, which can affect washing and, consequently, the purity of the extracted mRNA. In the case of bead aggregation, it is advisable to disperse the beads as much as possible during washing to improve the extraction efficiency. If bead aggregation occurs, it is recommended to reduce the total RNA amount in subsequent experiments.
• Due to the susceptibility of RNA degradation, it is recommended to use the extracted mRNA promptly for downstream experiments such as RT-PCR. If immediate use is not possible, store the mRNA at -80°C.
• This product is intended for scientific research by professionals only and should not be used for clinical diagnosis or treatment, food or drug purposes, or stored in ordinary residential areas.
• For your safety and health, please wear appropriate laboratory attire and disposable gloves while handling.

Storage

Store at 4°C, valid for one year. If the  Oligo (dT)25 magnetic beads are not used for an extended period, they can be stored at -20°C, which allows for longer-term storage. The entire kit can be stored at -20°C for an extended period of time.

Related: mRNA Purification Kit with Magnetic Beads

SBS Genetech is recognized as one of the global major leading industry players in mRNA Raw Material by third-party market researchers. For more details, please visit Top 5 mRNA Vaccine & Therapeutics Raw Material Market Companies in Global Market 2022.

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

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