Principle and classification of magnetic bead method nucleic acid automatic extraction instrument
Principle and classification of magnetic bead method nucleic acid automatic extraction instrumentMagnetic bead method nucleic acid automatic extraction instrument can generally be divided into magnetic rod method and suction method.
1.The suction method, also known as the pipetting method
It is to achieve the extraction and purification of nucleic acid through the transfer of liquid. Generally, the pipetting is completed by controlling the mechanical arm through the operating system.
The process and principles include:
1) Lysis: Add lysate to the sample, and by repeated pipetting, mixing and heating, the cells are broken to release nucleic acids;
2) Adsorption: Add magnetic beads to the sample lysate and mix thoroughly by blowing and beating. The magnetic beads have a strong affinity for nucleic acids under high salt and low pH to adsorb nucleic acids. Under the action of an external magnetic field, the magnetic beads and the solution Separation, use the tip to remove the liquid and discard it to the waste tank, and discard the tip;
3) Washing: Remove the external magnetic field, replace with a new tip and add washing buffer, repeatedly pipetting and mixing to remove impurities, and remove the liquid under the action of the external magnetic field;
4) Elution: Remove the external magnetic field, use a new tip to add elution buffer, and mix by repeated pipetting, and the bound nucleic acid is separated from the magnetic beads to obtain purified nucleic acid.
2.The magnetic rod method
that is, the separation and purification of nucleic acid is realized by the transfer of magnetic beads. Generally, the movement of the magnetic rod in the instrument is used to realize the transfer of the magnetic beads from the sample lysate/binding solution to the washing solution, and then to the eluent, so as to automatically Complete the whole process of nucleic acid isolation and purification.
The process and principle include the following steps:
1) Lysis adsorption: Add the sample to be treated in the lysis solution containing magnetic beads, mix well, lyse the cells (appropriate heating is helpful for cell lysis), and the released nucleic acids are specifically adsorbed to the magnetic beads under high salt and low pH values. on the beads, while molecules such as proteins are not adsorbed and remain in solution;
2) Washing: Under the action of the magnetic field of the magnetic bar, the magnetic beads are separated from the solution, and the magnetic bar transfers the magnetic beads to the washing buffer, and after repeated washing, impurities such as protein inorganic salts are removed;
3) Elution: After washing, the magnetic bar transfers the magnetic beads to the elution buffer, and the nucleic acid is eluted from the magnetic beads at a low salt and high pH value, and finally the magnetic bar removes the magnetic beads, that is, Complete nucleic acid extraction.
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