Nucleic acid extraction method

An extraction method and nucleic acid technology, applied in DNA preparation, recombinant DNA technology, etc., can solve the problems of manual transfer of nucleic acid eluent, many washing and elution steps, and intensified fragmentation of nucleic acid, so as to avoid human interference and pollution, nucleic acid The effect of simplifying the extraction process, improving accuracy and consistency

Inactive Publication Date: 2018-08-17
SHAOXING INGENIGEN BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The existing problems of the traditional magnetic rod extraction method are: 1. The purified nucleic acid eluate needs to be transferred manually, and manual operation errors are prone to occur during the transfer process, resulting in contamination by human factors and false positives
2. After salt washing, washing, and elution, there are many washing and elution steps, which take a long time, and the error rate will be greatly increased, especially when a large number of samples need to be processed in a short period of time, the cross-contamination is serious and the cost is high. More manpower, and the error rate will increase significantly
3. The multiple t

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Embodiment 1

[0161] Minimum Detection Limit Testing for DNA References

[0162] Use the nucleic acid extraction method of the present invention to manually operate the test, and use a nucleic acid extractor to automatically extract nucleic acid for testing to verify the feasibility:

[0163] Using the nucleic acid extraction method of the present invention to manually operate the experiment, the steps are as follows:

[0164] Step 1. Obtain a PCR reaction tube containing lysis solution and magnetic beads (8-row PCR tubes are used in this experiment, and the sequence of holes is A, B, C, D, E, F, G, H). Add samples to the first PCR reaction tube; add 10 2 CFU / ml, Bacillus pertussis (DNA reference), D4, E4, F4 wells were added with 10 3 CFU / ml, Bacillus pertussis (DNA reference); G4, H4 wells were added with 10 4 CFU / ml, Bacillus pertussis (DNA reference); add 10 to the A5 well of the second PCR reaction tube 4 CFU / ml, Bacillus pertussis (DNA reference), B5, C5, D5 wells were added with ...

Embodiment 2

[0181] Anti-pollution test of nucleic acid extractor

[0182] 2.1 The first set of anti-contamination tests for DNA reference

[0183] Using the nucleic acid extractor of the present invention to operate the experiment, the steps are as follows:

[0184] S1. Obtain a PCR reaction tube containing lysis solution and magnetic beads (8-row PCR tubes are used in this experiment, and the sequence of holes is A, B, C, D, E, F, G, H). Add samples, the samples are arranged as Figure 15 shown,

[0185] The A3 well position of the first PCR reaction tube is a DNA strong positive reference 10 6 CFU / m (Bacillus pertussis), B3 hole is a negative sample, C3 hole is a DNA strong positive reference 10 6 CFU / m (Bacillus pertussis), D3 hole is a negative sample, E3 hole is a DNA strong positive reference 10 6 CFU / m (Bacillus pertussis), F3 well is a negative sample, G3 well is a DNA strong positive reference 10 6 CFU / m (Bacillus pertussis), the H3 hole is a negative sample;

[0186] The ...

Embodiment 3

[0239] 3.1. Precision testing of DNA reference materials

[0240] Using the nucleic acid extractor of the present invention to operate the experiment, the steps are as follows:

[0241] S1. Obtain a PCR reaction tube containing lysis solution and magnetic beads (8-row PCR tubes are used in this experiment, and the sequence of holes is A, B, C, D, E, F, G, H). Add samples, the samples are arranged as Figure 15 As shown, prepare four 8-row PCR reaction tubes containing lysis buffer and magnetic beads, and add DNA reference material (10 6 CFU / ml, Bacillus pertussis);

[0242] Prepare 4 8-row PCR reaction tubes containing the pretreatment solution and place them in the tube rack in the pretreatment area;

[0243] S2. Put the four PCR reaction tubes after loading into the heating load of the lysis area, and put each hole in a sleeve of the heating load; liquid heating, the temperature rises, the exhaust fan is turned on, and after the heating is completed, the pyrolysis liquid...

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Abstract

The invention discloses a nucleic acid extraction method. The nucleic acid extraction method comprises the steps of cracking and transferring. In the cracking step, thermal cracking of nucleic acid iscompleted, and magnetic beads adsorb nucleic acid released in the thermal cracking process, and the cracking step is completed in a cracking area; the magnetic beads adsorbing the nucleic acid are transferred into pre-treatment fluid, and the transferring step is completed; and the pre-treatment fluid is located in a pre-treatment area, and the cracking area is mutually independent of the pre-treatment area. The nucleic acid extraction method has the advantages that the nucleic acid extraction steps are simplified, the extraction period is shortened, the nucleic acid extraction success rate is improved, and loss of the nucleic acid extraction amount is less.

Description

technical field [0001] The present invention relates to the field of nucleic acid extraction, more specifically, to a nucleic acid extraction method. Background technique [0002] The extraction of nucleic acid, such as DNA or RNA, currently mainly adopts two extraction methods: spin column method and magnetic bead method. The traditional spin column technology also uses membranes for nucleic acid purification. The principle of its membrane belongs to the silicon adsorption method. This membrane only has a strong affinity and adsorption force for nucleic acids. During the operation, the sample was firstly lysed by protease at a temperature of 56 °C, and then the lysing solution was added to the column for centrifugation, the nucleic acid was adsorbed on the membrane, and other impurities were thrown out, and finally the nucleic acid was eluted with the eluent. This method has good extraction effect, but requires heating during lysis, and multiple centrifugations during nucl...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 杨尚鑫胡彬刘杰蔡媛媛陶施芳
Owner SHAOXING INGENIGEN BIOTECH CO LTD
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