Nucleic acid extraction method and extraction agent thereof
An extraction method and reagent technology, applied in the field of biological detection methods and reagents, can solve the problems of easy cross-contamination between samples, cumbersome nucleic acid extraction steps, and contamination between samples, so as to avoid sample contamination, reduce steps and difficulties, and reduce reagents. cost effect
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Embodiment 1
[0038] Nucleic acid rapid extraction reagent preparation:
[0039] First dissolve Tris and EDTA with DEPC water, the concentration of Tris is 5-10mM; the concentration of EDTA is 5-10mM; adjust the pH to 6-9, add Tris and EDTA solution, add in 3-[(3-cholesteryl aminopropyl )Dimethylamino]-1-propanesulfonic acid, the volume percentage concentration is 0.005%-0.02%; adding NP-40, the percentage content concentration is 0.01%-0.2%; adding Tween-20, the volume percentage content The concentration is 0.01-0.2%; adding Chelex-100, the volume content is 1-20%.
Embodiment 2
[0041] Nucleic acid extraction and fluorescence quantitative PCR amplification of hand, foot and mouth virus throat swab
[0042] The step of nucleic acid extraction: get 12 routine pharyngeal swab samples to extract respectively with domestic magnetic bead method (CZ), imported silicon substrate membrane (GJ) and extraction reagent (KT) of the present invention,
[0043] Quick (KT) reagent extraction nucleic acid step of the present invention:
[0044] 1) Take 30 μl swab sample and add it to 50 μl extraction solution, lyse at 90°C for 4 minutes, and beat once evenly during the lysis period. Take it out and leave it at room temperature until the particles settle, and the supernatant is the obtained nucleic acid.
[0045] Steps for nucleic acid extraction with domestic magnetic beads (CZ) reagents:
[0046] 1) Take 200 μl of virus liquid into a 1.5ml centrifuge tube (prepared by yourself).
[0047]2) Add 15 μl of magnetic bead suspension G to the centrifuge tube.
[0048] 3...
Embodiment 3
[0077] Serum nucleic acid extraction and HBV fluorescence quantitative PCR amplification
[0078] The steps of nucleic acid extraction: 10 cases of serum samples were extracted by domestic magnetic bead method (CZ), imported silicon matrix membrane (GJ) and extraction reagent (KT) of the present invention.
[0079] Quick (KT) reagent extraction nucleic acid step of the present invention:
[0080] 1) Take 10 μl serum sample and add it to 50 μl extract solution, lyse at 90°C for 4 minutes, and beat evenly once during the lysis period. Take it out and leave it at room temperature until the particles settle, and the supernatant is the obtained nucleic acid.
[0081] The imported silicon matrix membrane (GJ) and the extraction reagent (KT) of the present invention are extracted according to the steps of Example 2.
[0082] HBV nucleic acid amplification was amplified with the "Hepatitis B Virus Nucleic Acid Detection Kit (Fluorescence PCR)" kit produced by Beijing Jinhao Pharmace...
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