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Kit for extracting dried blood spot genome DNA by paramagnetic particle method and extraction method

A kit and technology of dried blood spot, applied in the field of molecular biology, can solve the problems of incapability of using machines, general extraction efficiency, and inappropriate high-throughput operation, and achieve improved quality and total amount, high extraction efficiency, and good purity. Effect

Inactive Publication Date: 2021-07-16
GENFINE BIOTECH BEIJING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are two main methods for extracting genomic DNA from dried blood spots: column membrane method and magnetic bead method. It has been basically not used; in addition, column membrane extraction cannot be used for machines, nor is it suitable for automatic high-throughput operations, and the extraction efficiency is average
[0003] At present, it is difficult to extract the complete genomic DNA from dried blood spots, and it is difficult to achieve no fragmentation and smearing. Therefore, it is urgent to develop a dried blood spot genomic DNA Extraction reagents and kits to extract genomic DNA with complete fragments, stable quality, and neat bands, so as to meet the needs of corporate customers and relevant personnel in research institutes, major universities, and medical institutions

Method used

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  • Kit for extracting dried blood spot genome DNA by paramagnetic particle method and extraction method
  • Kit for extracting dried blood spot genome DNA by paramagnetic particle method and extraction method
  • Kit for extracting dried blood spot genome DNA by paramagnetic particle method and extraction method

Examples

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Effect test

Embodiment 1

[0061] Example 1. Using the magnetic bead method to extract genomic DNA from dried blood spots using a kit for extracting genomic DNA from dried blood spots.

[0062] In this example, the kit for extracting genomic DNA from dried blood spots by the magnetic bead method includes a lysate, a magnetic bead and a magnetic bead binding solution, a rinsing solution, an eluent, and proteinase K. In this kit, the lysis buffer MDA is: guanidine isothiocyanate 3mol / L, SDS 3% (w / v), Tris-HCl 60mmol / L, EDTA 5mmol / L, NaCl 10mmol / L, Triton X-1001.0 % (v / v), Tween-20 1.0% (v / v).

[0063] The magnetic bead binding solution Buffer DLTP is: 50mM Tris 50mM EDTA, 2% Triton X-100.

[0064] The first rinse solution is: Buffer DW: PEG 6000 6% (w / v) and NaCl 30mmol / L, and the solvent is sterile water.

[0065] The second rinse solution is: Buffer MWE2: 75% (v / v) ethanol.

[0066] Eluent Buffer EB: 1M Tris or RNase-free ddH at pH 8.0 2 O.

[0067] The magnetic beads are 300nm silanol magnetic bea...

Embodiment 2

[0079] Example 2, the screening of magnetic beads in the kit for extracting genomic DNA from dried blood spots by magnetic bead method

[0080] 1. Screening of magnetic bead types

[0081] In this embodiment, 14 different magnetic beads are selected, which are the first type: magnetic beads ABP; the second type: gold 1; the third type: gold 2; the fourth type: gold 3; the fifth type: gold 4; 6th type: gold 5; 7th type: FineMag G beads; 8th type: 1; 9th type: 2; 10th type: 3; 11th type: 4; 12th nanometer; No. 13: BSMOH-500; No. 14 BPOH-300. Among them, the 1st to 7th types are silicon hydroxyl magnetic beads; the 8th to 14th types are silicon carboxyl magnetic beads.

[0082] like figure 1 and figure 2 , the present embodiment adopts the magnetic bead method of embodiment 1 to extract the test results of the screening process of extracting genomic DNA from dried blood spot samples. In this embodiment, 14 kinds of magnetic beads have been screened, and finally the 7th magne...

Embodiment 3

[0088] Example 3, Screening of Magnetic Bead Binding Solution in Genomic DNA Extraction Kit for Screening Dried Blood Spot Samples

[0089] In this embodiment, the extraction method described in Example 1 is used to screen the sample magnetic bead binding solution, and the combination of magnetic beads and genomic DNA after cell lysis needs to be in a suitable magnetic bead binding solution. In order to increase the concentration of extracted DNA, this embodiment respectively Add an equal volume of isopropanol to seven magnetic bead binding solutions DLT, DLB, GHL, GHC, DP1, MLP1 and AL.

[0090] Determination of the extraction optimal conditions of the kit of the embodiment of the present invention on different human dried blood spot samples, such as Figure 5 As shown, in the dried blood spot samples of 5 people, the extraction efficiency of the genome of magnetic beads E and magnetic beads F under the conditions of different magnetic bead binding solutions DLB + isopropanol...

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Abstract

The invention discloses a kit for extracting dried blood spot genome DNA by a paramagnetic particle method and an extraction method. A DNA extraction reagent comprises a lysis solution, magnetic beads and a magnetic bead binding solution, wherein in the magnetic bead binding solution, the mass concentration of guanidine thiocyanate is 1-10 M / L, the mass concentration of Tris is 10-100 mM / L, the mass concentration of EDTA is 10-100 mM / L, and the volume fraction of Triton X-100 is 2%; or in the binding solution, the mass concentration of the guanidine thiocyanate is 1-10M / L, the mass concentration of the Tris is 10-100mM / L, the mass concentration of the EDTA is 10-100mM / L, and the volume fraction of the Tween 20 is 1.5%; and the magnetic beads are silicon hydroxyl magnetic beads. According to the dried blood spot genome nucleic acid kit and the extraction method disclosed by the invention, various blood-related samples, such as dried blood spots, blood samples, blood clot samples, blood gauze and tissue samples rich in blood cells, can be extracted; and the extraction efficiency is high, the purity is good, and the product stability is strong.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a kit and an extraction method for extracting genomic DNA from dried blood spots by a magnetic bead method. Background technique [0002] Extracting genomic nucleic acid from dried blood spots is a very common requirement, such as infant genetic disease screening, paternity testing, detection of multiple diseases in adults, forensic investigation, etc., it is necessary to extract the genome of lymphocytes in dried blood spots, through the The analysis of the genome is very important for the diagnosis and treatment of clinical medical diseases. At present, there are mainly two methods for extracting genomic DNA from dried blood spots: column membrane method and magnetic bead method. It has been basically not used; in addition, column membrane extraction cannot be used in machines, nor is it suitable for automatic high-throughput operations, and the extraction efficiency...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 韩典霖胡佳佳胡美丽杨亮
Owner GENFINE BIOTECH BEIJING CO LTD
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