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Rapid extraction method of oral mucosa cell genome DNA

A technology for oral mucosal cells and extraction methods, which is applied in the field of rapid extraction of genomic DNA from oral mucosal cells, can solve the problems of increased cost and extraction time, and achieve good application effects, simple ingredients, and strong pertinence

Pending Publication Date: 2019-02-15
南通中科医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] There are many extraction kits for genomic DNA of oral mucosal cells on the market, and the DNA extraction quality of most kits is maintained at a

Method used

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  • Rapid extraction method of oral mucosa cell genome DNA

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Oral mucosal cells from 12 individuals were collected using ordinary buccal swabs.

[0026] Follow the steps below to extract DNA:

[0027] 1) Add 450 μL of NaOH (50 mM) into the test tube, vortex and mix for 10 seconds, and place at 95°C for 5 minutes;

[0028] 2) Leave it at room temperature for 2 minutes and centrifuge immediately, take out the swab with tweezers, add 55 μL Tris-Hcl (20 mM) solution into the tube, and vortex for 10 seconds. Centrifuge at 12000 rpm for 1 min, and take 45 μL of supernatant.

[0029] 3) Add 90 μL of magnetic beads to the PCR tube containing the above supernatant, gently pipette and mix 6 times, incubate at room temperature for 5 minutes, and place the PCR tube on the magnetic stand for 3 minutes;

[0030] 4) Remove the supernatant, continue to place the PCR tube on the magnetic stand, add 300 μL of 75% ethanol solution to the PCR tube, rotate the tube 180 degrees twice, and let it stand for 30 seconds.

[0031] 5) Remove the supernat...

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Abstract

The invention provides a rapid extraction method of oral mucosa cell genome DNA. The method comprises the steps as follows: 1) adding 450-500 mu L of NaOH (50 mM) to a test tube, uniformly mixing NaOHfor 10-45 s through vortex, and leaving the mixture to stand at 95 DEG C for 5-6 min; 2) leaving the mixture to stand at the room temperature, performing instant centrifugation, taking out a swab with forceps, adding 55-60 mu L of a Tris-Hcl (20 mM) solution to the tube, uniformly mixing the mixture for 10-15 s through vortex, performing centrifugation at 12,000 rpm for 1 min, and taking 45-55 muL of a supernatant; 3) adding 90-95 mu L of magnetic beads to a PCR tube containing the supernatant, lightly sucking, beating and uniformly mixing the mixture 6 times, leaving the mixture to stand atthe room temperature for incubation for 5-10 min, and placing the PCR tube in a magnetic rack for 3-5 min. The method has the technical effects as follows: 1) only 4 reagents are involved in the whole DNA extraction process, components are simple, and the requirement for purity and the like is not high.

Description

technical field [0001] The invention relates to a method for rapidly extracting genomic DNA of oral mucosa cells, belonging to the field of biotechnology. Background technique [0002] There are many extraction kits for genomic DNA of oral mucosal cells on the market, and the DNA extraction quality of most kits is maintained at about 1.8 in OD260 / OD280. In order to increase the amount of DNA extraction, multiple steps may be involved, so the cost and extraction time will increase. [0003] With the continuous development of detection technology, the sensitivity of the instrument itself has increased, the cost is low, the operation is simple, the time-consuming is short, and the obtained DNA can conform to most platforms, which has become the first choice of most genetic testing industry. Contents of the invention [0004] The purpose of the present invention is to solve the problems existing in the prior art, and to provide a method for rapidly extracting genomic DNA of o...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 陆军姜昕刘慧君景丹丹
Owner 南通中科医学检验实验室有限公司