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Application of silica gel solution in extraction of human oral cavity cell DNA

A technology of oral cells and silica gel, applied in the field of DNA extraction, can solve the problems of inapplicable one-time processing of multiple samples, low DNA yield, and unusability

Active Publication Date: 2011-07-20
XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And its effect of obtaining DNA is better than the phenol-chloroform method, but due to its high cost, the amount of DNA extracted at one time is small, and cannot be used in large-scale production
[0006] Although there are some techniques that use membranes or resins (such as silica gel membranes) to absorb and separate DNA, this method is not suitable for occasions where the amount of biological samples is small, such as detecting oral samples of only about 1ml; and, this method Also not suitable for processing multiple samples at once
In addition, oral cells contain a variety of enzymes and degradable substances that are not found in other types of cell samples. The traditional method of extracting oral cell DNA often encounters the problem of low DNA yield or even complete degradation. At present, there is no specific method for oral DNA extraction products from cell samples

Method used

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  • Application of silica gel solution in extraction of human oral cavity cell DNA
  • Application of silica gel solution in extraction of human oral cavity cell DNA
  • Application of silica gel solution in extraction of human oral cavity cell DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Embodiment 1 obtains the method for DNA from oral cavity cell sample

[0094] The technological process of this embodiment sees figure 1 As shown, the specific steps are as follows:

[0095] (1) Obtain an oral cell sample (about 1.5ml) from the subject's oral cavity with a DNA sampling swab, and place the obtained sample in a sample tube;

[0096] (2) Add 600 μl of extraction buffer (Extraction Buffer), and put it in a 56°C warm bath to fully react;

[0097] (3) Add 1ml of lysis buffer (Lysis Buffer), 80μl of silica gel adsorbent (1.5mg) and mix well. and fully reversed;

[0098] (4) After sufficient centrifugation, discard the supernatant;

[0099] (5) 600 μl of washing buffer (Washing Buffer) resuspended;

[0100] (6) Discard the supernatant after sufficient centrifugation;

[0101] (7) Resuspend in 500 μl 70% ethanol twice and centrifuge fully;

[0102] (8) Resuspend in 500 μl acetone and centrifuge thoroughly;

[0103] (9) Dry at room temperature for 2-5 min...

Embodiment 2

[0112] The comparison of embodiment 2 silica gel adsorption method and phenol chloroform method, DNA extraction kit method

[0113] figure 2 , image 3 , Figure 4 In order to use the same amount of oral cell samples, DNA was extracted by the above three methods respectively. Among them, 1 and 2 are the silica gel adsorption method; 3 and 4 are the phenol-chloroform method; 5 and 6 are the Boguang DNA extraction kit method for extraction.

[0114] figure 2 It is the result of extraction electrophoresis without RNaseA digestion. It can be seen that before RNaseA digestion, 3, 4, 5, and 6 are mixed with a lot of RNA, and the effect is very poor.

[0115] image 3 It is the electrophoresis result after RNaseA treatment at 55°C for 30 minutes. It can be seen that the extraction results of 1 and 2 using silica gel adsorption method and 5 and 6 using DNA extraction kit method are significantly better than 3 and 4 phenol chloroform method, while the silica gel adsorption metho...

Embodiment 3

[0117] Embodiment 3 large-scale extraction

[0118] On a 96-well plate, 1.5 ml of buccal cell samples collected from different subjects were added to multiple wells. For the sample in each hole, the specific operation process is the same as in Example 1.

[0119] It can be seen that the silica gel adsorption method of the present invention can process multiple samples at one time, which is better than the DNA extraction kit.

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Abstract

The invention discloses a new rapid extracting method of DNA from oral cell, which comprises the following steps: (1) cracking pre-extracted DNA cell; obtaining cracking liquid of cell; (2) adding silicon gel particle into the cracking liquid; (3) separating the adsorbed DNA silicon gel particle; (4) obtaining DNA from silicon gel with low cost and high receiving rate of DNA; fitting for large-scale of DNA extraction.

Description

technical field [0001] The invention relates to the field of biotechnology, more specifically, the invention relates to a method for extracting DNA from a sample. Background technique [0002] At present, the technology of extracting DNA from cell samples has been more and more widely used, and the obtained DNA can be used for various detection or scientific research, for example. [0003] Extracting DNA from human oral cells is simpler, faster, non-invasive and clean than other methods, and is being more and more widely used. [0004] At present, there are mainly two methods for extracting DNA: phenol-chloroform method and DNA extraction kit method. The phenol-chloroform method is mainly used to extract DNA from human whole blood, and sometimes it is also used to extract DNA from human oral cells. Because of its low cost, it is widely used. The main disadvantage of this method is that saturated phenol and chloroform-isoamyl alcohol are added during the extraction process...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14
Inventor 邓新达闫鹏荣徐梦晔卢大儒
Owner XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI