Method and apparatus for concentrating and preserving genomic DNAs in urine sample

A urine sample, genome technology, applied in the biological and medical fields, can solve problems such as labor and material resources

Inactive Publication Date: 2018-07-31
BEIJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is very labor-intensive to preserve so many urine clinical samples in the form of liquid cryopreservation

Method used

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  • Method and apparatus for concentrating and preserving genomic DNAs in urine sample
  • Method and apparatus for concentrating and preserving genomic DNAs in urine sample
  • Method and apparatus for concentrating and preserving genomic DNAs in urine sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Concentration, collection and preservation of urine DNA

[0043] 1. Materials and Reagents

[0044] 1.1 Instrument: see device figure 1 .

[0045] 1.2 Materials and reagents: purified water; small tweezers; nylon membrane with a pore size of 0.22 μm; 102-type medium-speed qualitative filter paper; scissors; vacuum packaging machine; packaging bags.

[0046] 2. Experimental method

[0047] (1) Freshly collected urine samples from volunteers;

[0048] (2) Preparation of nylon membrane and filter paper:

[0049] Each urine sample (approximately 20ml to 50ml) is stored on one or more 0.22μm nylon membranes. Cut the corresponding number of circular nylon membranes and filter papers with a diameter of 47 mm as required.

[0050] Place 3 to 5 circular filter papers moistened with water on the Buchner funnel.

[0051] Wash the nylon membrane by immersing it in pure water, and then place it on a round filter paper, taking care to avoid the generation of air bub...

Embodiment 2

[0057] Example 2: Concentration, collection and preservation of urine DNA

[0058] 1. Materials and Reagents

[0059] 1.1 Instrument: see device figure 2 , wherein the membrane replacement filter 2 is composed of upper and lower parts, and the upper and lower parts have cylindrical holes that can be connected with other devices, and the upper cylindrical holes are connected with the syringe 1 (without needle) to provide pressure difference . The nylon membrane is located between the upper and lower parts of the membrane change filter.

[0060] 1.2 Materials and reagents: purified water; small tweezers; nylon membrane with a pore size of 0.22 μm; 102-type medium-speed qualitative filter paper; scissors; vacuum packaging machine; packaging bags.

[0061] 2. Experimental method

[0062] (1) Freshly collected urine samples from volunteers;

[0063] (2) Preparation of nylon membrane:

[0064] Each urine sample (approximately 20ml to 50ml) is stored on one or more 0.22μm nylo...

Embodiment 3

[0087] The extraction of the DNA preserved on the nylon membrane of embodiment 3

[0088] (1) Shred the nylon membrane, put it in a centrifuge tube, add 1.2ml of lysate, and vortex for 1min.

[0089] (2) Place in a water bath at 60°C for 2 hours, or overnight.

[0090] (3) Add 500 μl of phenol:chloroform (1:1), and vortex for 30 seconds.

[0091] (4) Centrifuge at 12,000 rpm for 15 minutes at 4°C.

[0092] (5) Take the supernatant, add 500 μl phenol:chloroform (1:1), shake and mix.

[0093] (6) Centrifuge at 12,000 rpm for 10 min at 4°C.

[0094] (7) Take the supernatant, add an equal volume of isopropanol, and mix well.

[0095] (8) Centrifuge at 12,000 rpm for 15 min at 4°C.

[0096] (9) After discarding the supernatant, add 1 ml of 75% ethanol, shake and mix well, and centrifuge at 12,000 rpm for 10 min at 4°C.

[0097] (10) After discarding the supernatant, dry it in the air for 10-15 minutes. Be careful not to completely dry the precipitate, otherwise it will increa...

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Abstract

The invention relates to a method and apparatus for concentrating and preserving genomic DNAs in a urine sample. Specifically, the invention relates to a method and apparatus for concentrating and preserving DNAs in urine by using a membrane capable of adsorbing DNAs. According to the method, DNAs in urine are adsorbed onto the membrane and preserved in vacuum in a dried form; the method is simpleand easily practicable, has low cost, does not need any organic solvent and causes no pollution to environment; and the method can concentrate low-concentration DNAs in urine and preserve the DNAs under complete vacuum and dry conditions, so the degradation of the DNAs is avoided, the DNAs in urine can be stored at room temperature for a long time, the DNAs of each component in urine are retained, and a foundation is laid for subsequent construction of a biological sample library with more comprehensive information.

Description

technical field [0001] The invention relates to the fields of biology and medicine, in particular to a method for concentrating, collecting and preserving genomic DNA in a biological sample and a device thereof. Background technique [0002] In the late 1990s, scientists proposed the concept of biological sample bank, which is to standardize the collection, processing, storage and application of samples of biological macromolecules, cells, tissues and organs of healthy and diseased organisms, including human organ tissues, whole blood, Plasma, serum, biological fluid or processed biological samples, and biological application systems for clinical treatment of diseases and life science research. [0003] Urine is a waste product excreted by the body, and it can be obtained in large quantities non-invasively and continuously. More and more studies have gradually proved that urine can not only dynamically reflect changes in the kidney and urinary system, but also reflect abnor...

Claims

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Application Information

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IPC IPC(8): G01N1/40B65D81/20
CPCB65D81/2023G01N1/405
Inventor 高友鹤张繁霜倪艳颖
Owner BEIJING NORMAL UNIVERSITY
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