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Kit for extracting genome DNA of oral sample based on magnetic bead method and application method of kit

A genomic and magnetic bead method, applied in the field of kits, can solve the problems that magnetic beads cannot efficiently capture nucleic acids, weak ability to capture nucleic acids, and small specific surface area of ​​magnetic beads. Efficient effect

Inactive Publication Date: 2017-08-18
SUZHOU ENRICHING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing magnetic bead method still cannot achieve the technical problems of no need for centrifugation and high-throughput quantification, and the magnetic beads used in the prior art have a small specific surface area and a high density, resulting in poor suspension ability and poor ability to capture nucleic acids. Weak, when the amount of nucleic acid in the sample is small, the magnetic beads cannot efficiently capture nucleic acid

Method used

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  • Kit for extracting genome DNA of oral sample based on magnetic bead method and application method of kit
  • Kit for extracting genome DNA of oral sample based on magnetic bead method and application method of kit
  • Kit for extracting genome DNA of oral sample based on magnetic bead method and application method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] A kit for extracting genomic DNA from oral samples based on the magnetic bead method, comprising:

[0044] Preservation solution: 7mmol / L disodium edetate, 15mmol / L Tris alkali aqueous solution, pH value is 7.0. Specific preparation method: Weigh 1.82g Tris base and 2.61g disodium edetate, add 800mL deionized water to fully dissolve, continue to add deionized water to make the volume to 1L, and adjust the pH value to 7.0 with concentrated hydrochloric acid.

[0045] Proteinase K solution: 22g / L proteinase K solution. Specific preparation method: Weigh 2.20g of proteinase K, add 80mL of deionized water to dissolve, continue to add deionized water to make up to 100mL.

[0046]Lysis solution: 6mol / L of guanidine hydrochloride, 0.3mol / L of sodium chloride, 2% (w / v) of polyvinylpyrrolidone, 2% (w / v) of cetyltrimethylammonium bromide and 7mmol / L mixed solution of disodium edetate. Specific preparation method: Weigh 573.18g of guanidine hydrochloride, 17.53g of sodium chlor...

Embodiment 2

[0070] A kit for extracting genomic DNA from oral samples based on the magnetic bead method, comprising:

[0071] Preservation solution: 3mmol / L dipotassium edetate, 5mmol / L Tris alkali aqueous solution, pH value is 7.0. Specific preparation method: Weigh 0.61g of Tris base and 1.12g of dipotassium edetate, add 800mL of deionized water to fully dissolve, continue to add deionized water to adjust the volume to 1L, and adjust the pH value to 7.0 with concentrated hydrochloric acid.

[0072] Proteinase K solution: 18g / L proteinase K solution. Specific preparation method: Weigh 1.80g proteinase K, add 80mL deionized water to dissolve, continue to add deionized water to make up to 100mL.

[0073] Lysis solution: guanidine isothiocyanate of 1mol / L, sodium chloride of 0.05mol / L, polyvinylpyrrolidone of 0.5% (w / v), hexadecyl trimethyl bromide of 0.5% (w / v) A mixed solution of ammonium chloride and 3 mmol / L dipotassium edetate. Specific preparation method: Weigh 118.16g of guanidine...

Embodiment 3

[0092] Use the kit of Example 1 to carry out high-throughput extraction experiments on 27 parts of saliva genomic DNA:

[0093]Step 1: Add 200 μL saliva sample, 20 μL proteinase K solution and 300 μL lysate to the 48-well plate in sequence with a multi-channel automatic pipette, and 27 samples in parallel, vortexed to mix;

[0094] Step 2: Place the above sample in a warm bath at 58°C for 15 minutes, and shake it gently several times during this period;

[0095] Step 3: Take out the 48-well plate, add 400 μL of isopropanol and 100 μL of magnetic bead dispersion to the 27 sample wells with a multi-channel automatic pipette, place the 48-well plate on a vortex mixer, and incubate with high-speed vortex mixing 5min;

[0096] Step 4: Place the 48-well plate on the magnetic stand and let it stand for 30-90 seconds. After the magnetic beads are completely absorbed, pour and discard the supernatant, and then place it upside down on absorbent paper to completely remove the residual s...

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Abstract

The invention discloses a kit for extracting genome DNA of an oral sample based on a magnetic bead method and an application method of the kit. The kit comprises a preservation liquid, a protease K solution, a lysate, a magnetic bead dispersion liquid, a cleaning liquid I, a cleaning liquid II and an eluent. The kit is simple and convenient to operate, and the work efficiency of extracting saliva or swab genome DNA can be greatly improved; and saliva does not need to be pretreated, and a saliva or swab sample can be split by adopting the lysate directly, and the magnetic bead dispersion liquid is added to be combined with the genome DNA, so that the extraction yield of nucleic acid is high and the kit is suitable for a high-throughput experiment.

Description

technical field [0001] The invention relates to a kit, in particular to a kit for extracting genomic DNA from an oral cavity sample based on a magnetic bead method and a use method thereof. Background technique [0002] In recent years, with the rapid development of molecular biology, researchers have paid more and more attention to DNA. Whether it is gene sequencing, PCR amplification, or construction of BCA library, etc., there are high requirements on the concentration, purity, and integrity of the primary structure of DNA, so high-quality DNA and a large number of DNA samples need to be extracted. But genomic DNA samples have traditionally been taken from blood. This blood collection method will cause certain harm and pain to the person being collected, and is disgusted or rejected by most people. Since saliva and blood have a certain similarity, and saliva contains a large number of exfoliated oral epithelial cells, collecting oral samples by collecting saliva or oral...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 吴巧宗洁尚春庆曲峰
Owner SUZHOU ENRICHING BIOTECH CO LTD
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