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Nucleic acid extractor

A nucleic acid extraction instrument and regional technology, applied in the field of nucleic acid extraction, can solve the problems of multiple washing and elution steps, intensified nucleic acid fragmentation, unfavorable treatment, etc., to simplify the nucleic acid extraction process, improve accuracy and consistency, and avoid interference and pollution Effect

Pending Publication Date: 2018-08-24
SHAOXING INGENIGEN BIOTECH CO LTD
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  • Summary
  • 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 transfers of magnetic beads lead to inevitable damage to the nucleic acid content. This is because the magnetic beads are exposed, and the nucleic acids adsorbed on the magnetic beads are exposed to the outside, and are susceptible to impurities in the environment, such as dust, enzymes, or other substances. Material contamination, especially for RNA, is more susceptible to degradation
4. Nucleic acid undergoes repeated washing, elution and heating, which may significantly reduce the integrity of nucleic acid and intensify the fragmentation of nucleic acid, which is not conducive to subsequent processing, such as subsequent sequencing, amplification or other utilization

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0167] Minimum detection limit test for DNA reference

[0168] Use the nucleic acid extraction method of the present invention to manually operate the test, and use the nucleic acid extractor to automatically extract nucleic acid for test verification feasibility:

[0169] Using the nucleic acid extraction method of the present invention to manually operate the experiment, the steps are as follows:

[0170] Step 1. Obtain a PCR reaction tube containing lysate and magnetic beads (this test uses 8 rows of PCR tubes, and the order of the holes is A, B, C, D, E, F, G, H), and insert the PCR reaction tube Add samples; add 10 2CFU / ml, Bacillus pertussis (DNA reference product), D4, E4, F4 wells add 10 3 CFU / ml, Bacillus pertussis (DNA reference); G4, H4 wells add 10 4 CFU / ml, Bacillus pertussis (DNA reference product); add 10 to well A5 of the second PCR reaction tube 4 CFU / ml, Bacillus pertussis (DNA reference product), add 10 5 CFU / ml, Bacillus pertussis (DNA reference produc...

Embodiment 2

[0187] Anti-pollution test of nucleic acid extractor

[0188] 2.1 The first set of anti-pollution tests for DNA references

[0189] Use the nucleic acid extractor operation experiment of the present invention, the steps are as follows:

[0190] S1. Obtain a PCR reaction tube containing lysate and magnetic beads (this test uses 8 rows of PCR tubes, the order of the holes is A, B, C, D, E, F, G, H), and insert the PCR tube into the PCR reaction tube. Adding samples, the arrangement of samples is as follows Figure 15 as shown,

[0191] Well A3 of the first PCR reaction tube is DNA strong positive reference product 10 6 CFU / m (pertussis bacilli), B3 hole is negative sample, C3 hole is DNA strong positive reference sample 10 6 CFU / m (pertussis bacilli), D3 hole is negative sample, E3 hole is DNA strong positive reference sample 10 6 CFU / m (pertussis bacilli), F3 hole is negative sample, G3 hole is DNA strong positive reference sample 10 6 CFU / m (pertussis bacilli), H3 hole i...

Embodiment 3

[0245] 3.1. Precision test of DNA reference product

[0246] Use the nucleic acid extractor operation experiment of the present invention, the steps are as follows:

[0247] S1. Obtain a PCR reaction tube containing lysate and magnetic beads (this test uses 8 rows of PCR tubes, the order of the holes is A, B, C, D, E, F, G, H), and insert the PCR tube into the PCR reaction tube. Adding samples, the arrangement of samples is as follows Figure 15 As shown, prepare 4 8-row PCR reaction tubes filled with lysate and magnetic beads, and add DNA reference products (10 6 CFU / ml, B. pertussis);

[0248] Prepare 4 8-row PCR reaction tubes filled with pretreatment solution and place them in the tube rack in the pretreatment area;

[0249] S2. Put the four PCR reaction tubes after adding the sample into the heating load in the lysis area, and put each hole into a sleeve of the heating load; The liquid is heated, the temperature rises, the exhaust fan is turned on, and after the tempe...

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Abstract

The invention discloses a nucleic acid extractor which is provided with an operation space. A magnetic rod-magnetic rod sleeve module and a driving transmission mechanism are arranged in the operationspace, the driving transmission mechanism enables the magnetic rod-magnetic rod sleeve module to reciprocate or move, a cracking area and a pretreatment area are included in the operation space and are independent from each other, and the magnetic rod-magnetic rod sleeve module reciprocates in the cracking area and the pretreatment area. The nucleic acid extractor is simpler in extraction step, short in extraction time, higher in nucleic acid extraction rate and has little nucleic acid extraction loss.

Description

technical field [0001] The invention relates to the field of nucleic acid extraction, more specifically, it relates to a nucleic acid extraction instrument and an extraction method. Background technique [0002] Nucleic acid, such as DNA or RNA, is currently extracted by two methods: spin column method and magnetic bead method. Traditional spin column technology also uses membranes for nucleic acid purification. The principle of its membranes belongs to the silica adsorption method. This membrane only has strong affinity and adsorption for nucleic acids. During the operation, the sample is firstly lysed with protease at a temperature of 56°C, then the lysate is added to the column for centrifugation, the nucleic acid is adsorbed on the membrane, while other impurities are thrown off, and finally the nucleic acid is eluted with the eluent. This method has a good extraction effect, but heating is required for lysis, and multiple centrifugations are required for nucleic acid p...

Claims

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Application Information

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IPC IPC(8): C12M1/42C12M1/00C12N15/10
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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