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Plasma free nucleic acid extraction kit and application thereof

A technology of free nucleic acid and kits, which is applied in DNA preparation, microbial measurement/testing, recombinant DNA technology, etc. It can solve the problems of large sample demand, inconvenient high-throughput, automatic operation, and extraction methods that need to be improved.

Active Publication Date: 2018-02-06
重庆华大医学检验所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method uses a special silica-based adsorption material, which is characterized by: in the presence of high hydrochloric acid buffer, DNA can be specifically adsorbed, and impurities that cannot be combined with RNA and protein can be removed, while low-salt alkaline buffer can elute and bind to the adsorption column. However, the disadvantage of this method is that it requires a large amount of samples and consumes more samples. Its application to some rare samples is greatly limited. At the same time, the extraction of nucleic acids by spin column method requires repeated centrifugation, which is not convenient for high-throughput, Automated operation, especially in the fields of genetic diagnosis, monitoring and control of sudden outbreaks, and the use of spin column method to extract nucleic acid requires a large number of operators and equipment to meet the demand
[0004] Therefore, the current nucleic acid extraction, especially the extraction method of plasma free nucleic acid, still needs to be improved.

Method used

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  • Plasma free nucleic acid extraction kit and application thereof
  • Plasma free nucleic acid extraction kit and application thereof
  • Plasma free nucleic acid extraction kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0123] The inventor produces plasma free nucleic acid extraction kit of the present invention according to the following methods, specifically as follows:

[0124] One, enumerate the part reagent, instrument that adopts in the present embodiment below:

[0125] Kit production raw material supply

[0126] The raw materials for preparing each reagent in the kit of the present invention are provided by well-known overseas biological consumables manufacturers.

[0127] The main materials and suppliers are as follows:

[0128]

[0129]

[0130] Kit production conditions and production capacity

[0131] The production of this product needs to be prepared in a clean room (class 100,000) or completed in an ultra-clean workbench.

[0132] The product is currently in a small-scale production period, and the specific output needs to be produced according to market demand.

[0133] Kit production process requires equipment and consumables

[0134] Equipment required for product...

Embodiment 2

[0200] Prepare the plasma free nucleic acid extraction kit of the present invention according to the method of Example 1, the only difference is that the composition and formula of the kit are different, specifically, the kit of the present embodiment has a good formula as follows:

[0201] (1) Magnetic bead lysis binding solution, including:

[0202] 130mM Tris-HCl buffer at pH 8.3;

[0203] 128mM EDTA buffer;

[0204] 9.2% (w / v) SDS buffer;

[0205] 3.0% (w / v) sodium citrate buffer;

[0206] 6% (w / v) polyethylene glycol solution;

[0207] 4.5M solution of guanidine isothiocyanate; and

[0208] the remainder of the water,

[0209] Wherein, before use, 10% (w / v) isopropanol needs to be added to the magnetic bead lysis binding solution,

[0210] (2) The first cleaning solution, including:

[0211] 6.5M guanidine isothiocyanate solution;

[0212] 90mM Tris-HCl buffer at pH 8.2;

[0213] 60mM EDTA buffer; and

[0214] the remainder of the water,

[0215] Wherein, befor...

Embodiment 3

[0227] Prepare the plasma free nucleic acid extraction kit of the present invention according to the method of Example 1, the only difference is that the composition and formula of the kit are different, specifically, the kit of the present embodiment has a good formula as follows:

[0228] (1) Magnetic bead lysis binding solution, including:

[0229] 140mM Tris-HCl buffer at pH 8.4;

[0230] 138mM EDTA buffer;

[0231] 7.2% (w / v) SDS buffer;

[0232] 3.5% (w / v) sodium citrate buffer;

[0233] 7% (w / v) polyethylene glycol solution;

[0234] 4.8 M guanidine isothiocyanate solution; and

[0235] the remainder of the water,

[0236] Wherein, before use, 20% (w / v) isopropanol needs to be added to the magnetic bead lysis binding solution,

[0237] (2) The first cleaning solution, including:

[0238] 7M guanidine isothiocyanate solution;

[0239] 85mM Tris-HCl buffer at pH 8.2;

[0240] 64mM EDTA buffer; and

[0241] the remainder of the water,

[0242] Wherein, before us...

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Abstract

The invention discloses a plasma free nucleic acid extraction kit and application thereof, wherein the kit comprises (1) magnetic bead pyrolysis combination solution; (2) a first cleaning solution; (3) a second cleaning solution; (4) an elution solution. By using the plasma free nucleic acid extraction kit provided by the invention, the free desoxyribonucleic acid in the plasma can be fast extracted; in addition, the use is fast, simple and convenient; the centrifugation is not needed; once mass extraction only needs half an hour; the obtained nucleic acid has high quality; when the obtained nucleic acid is used for subsequent sequencing and detection, the result is accurate and reliable; the complete application to the existing noninvasive prenatal genetic testing technology can be realized; the important significance is realized in aspects of scientific study and clinic detection.

Description

technical field [0001] The invention relates to a plasma free nucleic acid extraction kit and application thereof. Background technique [0002] At present, the precipitation and centrifugation operations involved in the existing traditional nucleic acid extraction technology require the use of a large number of biological samples, and the steps of the traditional extraction technology are relatively complicated, time-consuming, and the yield is low. It is difficult to realize automatic operation, and most of the methods are still Operators need to be directly exposed to toxic chemical reagents. Therefore, with the rapid development of molecular biology and polymer materials, the traditional method of separating and extracting nucleic acids from liquid systems is gradually being replaced by new methods based on solid-phase adsorbent carriers. method replaced. [0003] Such emerging nucleic acid separation and extraction methods mainly include: rotating spin column extractio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013C12Q2523/308C12Q2563/143C12Q2563/149
Inventor 萧哲吴宗泽蔡贤杵徐快黄健申丹赵振东刘娜
Owner 重庆华大医学检验所有限公司
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