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Kit for extracting DNA (deoxyribonucleic acid) from white cells in human whole blood

A kit, white blood cell technology, applied in DNA preparation, recombinant DNA technology, microbial determination/inspection, etc., to reduce cross-contamination between tubes, avoid DNA loss, and shorten extraction time.

Active Publication Date: 2014-04-09
SANSURE BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a human whole blood DNA rapid extraction kit and its method for extracting DNA from human whole blood leukocytes, to solve the defects of existing whole blood nucleic acid extraction kits, to provide a fast and simple operation, high extraction efficiency Kit for rapid extraction of DNA from human whole blood leukocytes with high efficiency, safe use and low manufacturing cost

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] 1. Kit for DNA extraction from human whole blood leukocytes and preparation method of reagents therein:

[0019] ①Preparation of erythrocyte lysate: first add a small amount of sterile water to the volumetric flask, weigh and add sodium chloride so that the final concentration is 5mmol / L, add triton solution with a final volume ratio of 1%, and add the final concentration Glucose is 320mmol / L, add trishydroxymethylaminomethane with a final concentration of 0.01mol / L; add sterile water to the required volume, mix well to dissolve, use sodium hydroxide or hydrochloric acid solution to adjust the pH value to 8.2; Autoclave for 10 minutes.

[0020] ②Preparation of DNA extraction solution: first add a small amount of sterile water to a volumetric flask, weigh and add potassium chloride to a final concentration of 0.8mol / L, add ethylenediaminetetraacetic acid with a final concentration of 0.6mol / L, and add The final concentration is 1% sodium lauryl sulfate (mass volume rati...

Embodiment 2

[0027] 1. Kit for DNA extraction from human whole blood leukocytes and preparation method of reagents therein:

[0028] ① Preparation of erythrocyte lysate: same as in Example 1.

[0029] ②Preparation of DNA extraction solution: first add a small amount of sterile water to the volumetric flask, weigh and add potassium chloride to a final concentration of 0.1mol / L, add surfactin to a final concentration of 0.2mmol / L, Add sodium dodecylsulfonate (mass volume ratio) with a final concentration of 0.5% and ethanol with a final concentration of 0.2%; add sterile water to the required volume, mix well and dissolve, use sodium hydroxide or Adjust the pH to 8.2 with hydrochloric acid solution; autoclave for 15 minutes.

[0030] 2. Application of kit:

[0031] The application object of the kit is that the DNA concentration of Epstein-Barr virus is 1×10 4 IU of whole blood samples. The specific operation steps of the application are: add 800 microliters of red blood cell lysate to 20...

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Abstract

The invention provides a kit for extracting DNA (deoxyribonucleic acid) from white cells in human whole blood. The kit comprises a red blood cell lysis buffer and a DNA extracting solution, wherein the red blood cell lysis buffer comprises the following components: 4.5-5.5mmol / L sodium chloride, 0.8-1.2 percent (v / v) triton 100, 300-340mmol / L glucose and 0.008-0.012mol / L tri-hydroxymethyl aminomethane. The kit provided by the invention can be used for solving the defect of an existing kit for extracting the DNA in human whole blood white cells. The invention provides a kit for quickly extracting the DNA from white cells in the human whole blood, and the kit is simple and quick in operation, high in extraction efficiency, safe to use, and low in manufacture cost. By applying the kit, the DNA in white cells of a clinical whole blood sample for PCR (Polymerase Chain Reaction) technology detection can be rapidly extracted, and DNA in white cells and DNA of viruses in the white cells in whole blood are extracted.

Description

technical field [0001] The invention relates to a kit for extracting DNA from human whole blood leukocytes. Background technique [0002] Since the advent of PCR technology in the 1980s, its application in molecular biology-related fields has become more and more extensive. Especially in recent years, with the improvement of people's requirements for the accuracy and sensitivity of clinical diagnosis, and the introduction and promotion of the concept of individualized medicine, PCR technology has been rapidly and widely used in clinical diagnosis due to its advantages of rapidity, sensitivity and specificity in clinical diagnosis. Diagnosis and treatment, by detecting genes in patient samples to help doctors determine medication and treatment options. [0003] Sample DNA extraction is the prerequisite for the application of PCR technology. Human peripheral blood plays an important role in the immune response and metabolism in the human body, and can be used for molecular mo...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/68
Inventor 戴立忠唐瑶吴康邓中平
Owner SANSURE BIOTECH INC
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