Method for extracting DNA with pleuroperitoneal fluid tissue and kit

A kit, chest and abdomen technology, applied in the direction of recombinant DNA technology, DNA preparation, etc., can solve the problems of DNA quantity and purity not meeting the requirements of experimental analysis, incapable of large-volume batch operation, low extraction efficiency, etc., to achieve concentration and purity Good, meet the sample quality requirements, and easy to operate

Inactive Publication Date: 2019-05-14
杭州瑞普基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The composition of the pleural and ascites fluid is relatively complex. Extracting high-quality DNA from human pleural and ascites tissue is a prerequisite for routine molecular biology research and clinical related testing. There is no current DNA extraction method specifically for the use of pleural and ascites fluid extraction. DNA, and cannot be operated in large batches, so the extraction efficiency is low, and the amount and purity of the extracted DNA cannot meet the requirements of experimental analysis

Method used

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Examples

Experimental program
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Embodiment 1

[0044] The present embodiment provides a method for extracting DNA from pleural and ascites tissue, comprising the following steps:

[0045] 1) Take 10ml of pleural and ascites tissue samples under aseptic conditions, and place them in a 50ml sterile antifreeze centrifuge tube. Add 0.008 times the volume of 20% EDTA anticoagulant to the tube, and mix well by inverting up and down.

[0046] 2) Place the sample in step 1) in a centrifuge at 4° C. and centrifuge at 3000 g for 10 min to separate the supernatant of the pleural and ascites fluid from the pleural and ascites fluid precipitation.

[0047] 3) Discard the supernatant in step 2), and keep the precipitate. And 200ul of pH 8.0 PBS solution was added thereto, shaken and mixed.

[0048]4) Transfer the mixed solution in step 3) to a 1.5ml centrifuge tube, add 25ul 20mg / mL proteinase K, 200ul lysate, and vortex for 15s. Wherein, the concentration of each solvent component in the lysate is: 10mmol / L pH8.0 Tris-HCl, 400mmol / L...

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PUM

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a method for extracting DNA with pleuroperitoneal fluid tissue and a kit. The method for extracting DNA with pleuroperitoneal fluid tissue includes the following steps of pleuroperitoneal fluid pretreatment, cell lysis, nucleic acid adsorption, magnetic separation, protein removal and nucleic acid release from magnetic beads. The kit is prepared through the method. By the adoption of the method and the kit, the high-quality DNA can be efficiently and rapidly obtained, the demands, including PCR amplification,quantitative and qualitative analysis, sequencing and others, of molecular biology experiments can be met, application value is realized for diagnosis of related diseases generated by pleuroperitonealfluid, and a new conception is provided for preparation of DNA in pleuroperitoneal fluid tissue.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method and a kit for extracting DNA from pleural and ascites tissues. Background technique [0002] Pleural effusion is a type of localized edema, which is the accumulation of excessive fluid in the chest and abdominal cavity. The mechanism of pleural effusion is complex, which is related to the imbalance of fluid exchange in vivo and in vitro and the imbalance of fluid exchange in and out of blood vessels. A variety of malignant tumors can have pleural effusion, and the pleural effusion that occurs on the basis of tumors is called malignant pleural effusion. Therefore, pleural effusion is often used as one of the sources of tissue materials for disease diagnosis. [0003] The composition of the pleural and ascites fluid is relatively complex. Extracting high-quality DNA from human pleural and ascites tissue is a prerequisite for routine molecular biology research and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 臧德山
Owner 杭州瑞普基因科技有限公司
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