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Polystyrene magnetic bead based dried blood spot genome extraction kit and extraction method

A technology of polystyrene and genomics, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of sample cross-contamination, sample processing loss, multiple instruments, etc., and achieve high binding efficiency , not easy to settle, good uniformity

Inactive Publication Date: 2019-03-01
GUANGDONG ASCENDAS GENOMICS TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are a variety of genomic DNA extraction kits that can be used in various fields, but the traditional extraction technology requires precipitation, centrifugation and other operations, and requires a large number of biological samples, resulting in many shortcomings in genomic DNA extraction, such as the extraction process is complicated, the sample Long processing time, loss of a large amount of DNA in sample processing, error-prone extraction steps, easy to cause cross-contamination of samples, and the need for multiple instruments, etc.

Method used

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  • Polystyrene magnetic bead based dried blood spot genome extraction kit and extraction method
  • Polystyrene magnetic bead based dried blood spot genome extraction kit and extraction method
  • Polystyrene magnetic bead based dried blood spot genome extraction kit and extraction method

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Embodiment 1

[0029] Composition of the kit

[0030] The kit in this embodiment includes the following components: polystyrene magnetic beads, lysis solution, binding solution, washing solution and nucleic acid eluting solution. The polystyrene magnetic beads are hydroxyl magnetic beads coated with polystyrene gamma-ferric oxide.

[0031] The lysis solution includes a lysis solution I and a lysis solution II, and the lysis solution I includes tris, ethylenediaminetetraacetic acid, sodium chloride, Triton X-100, sodium dodecylsulfonate , the concentration of Tris is 20-80mmol / L, the concentration of EDTA is 5-20mmol / L, the concentration of sodium chloride is 50-500mmol / L, the concentration of Triton X-100 The concentration of sodium dodecylsulfonate is 1-5% by volume.

[0032] The lysate II comprises guanidine hydrochloride, ethylenediaminetetraacetic acid, sodium chloride, Triton X-100, sodium dodecylsulfonate, the concentration of guanidine hydrochloride is 1-8mol / L, trishydroxymethylami...

Embodiment 2

[0038] The method for extracting the dried blood slice genome in this embodiment is as follows:

[0039] 1) Use a puncher to punch 1-2 5mm diameter blood spots on the FTA Card or blood spots, and transfer them to a 2.0 ml centrifuge tube.

[0040] 2) Add 20 μL Proteinase K and 200 μL Lysis Solution I to the sample, and incubate at 60° C. for 20 minutes.

[0041] 3) Add Lysis Buffer II, invert and mix 5-10 times, and vortex at high speed for 20 seconds. Incubate at 70°C for 10 minutes.

[0042] 4) Transfer 350-400 μL of the lysate supernatant to a 1.5 ml centrifuge tube.

[0043] 5) Add 20ul polystyrene magnetic beads and 400μL binding solution, mix by inversion.

[0044] 6) Transfer to a magnetic stand for adsorption for 3-5 minutes, then discard the solution.

[0045] 7) Add 500 μL of washing solution I, and vortex to resuspend the magnetic beads. Transfer to a magnetic stand, absorb for 3-5 minutes, and discard the solution.

[0046] 8) Add 500 μL of washing solution II,...

Embodiment 3

[0052] The method for extracting the dried blood slice genome in this embodiment is as follows:

[0053] 1) Use a puncher to punch 1-2 5mm diameter blood spots on the FTA Card or blood spots, and transfer them to a 2.0 ml centrifuge tube.

[0054] 2) Add 20 μL Proteinase K and 200 μL Lysis Solution I to the sample, and incubate at 60° C. for 20 minutes.

[0055] 3) Add Lysis Buffer II, invert and mix 5-10 times, and vortex at high speed for 20 seconds. Incubate at 70°C for 10 minutes.

[0056] 4) Transfer 350-400 μL of lysed supernatant to row 1 / 7 of a 96-well plate, and add 20 μL of polystyrene magnetic beads.

[0057] 5) Add 700 μL of washing solution I to row 2 / 8 of 96 wells.

[0058] 6) Add 700 μL of washing solution II to rows 3 / 9 and 4 / 10 of the 96 wells.

[0059] 7) Add 50 μL of eluent to row 5 / 11 of 96 wells.

[0060] 8) Run the nucleic acid extractor, and transfer the 5 / 11 eluate to a 1.5ml centrifuge tube after completion.

[0061] Using the above method to ext...

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Abstract

The invention relates to a polystyrene magnetic bead based dried blood spot genome DNA extraction kit. The kit comprises polystyrene magnetic beads, splitting liquid, combination liquid, washing liquid and nucleic acid eluting liquid. The polystyrene magnetic beads are polystyrene coated gamma-iron trioxide hydroxyl magnetic beads. The invention further provides a dried blood spot genome DNA extraction method in which the kit is adopted for extraction. The method includes steps: S1, splitting dried blood spots by protease K and the splitting liquid, and taking supernatant to obtain a splittingproduct; S2, adding the polystyrene magnetic beads and the combination liquid into the splitting product, well mixing, transferring to a magnetic frame, and discarding solution; S3, adopting the washing liquid for washing; S4, adopting the eluting liquid for DNA eluting from the magnetic beads with DNA, so that genome DNA is obtained.

Description

technical field [0001] The invention relates to the field of blood sample detection, more particularly, to a polystyrene magnetic bead-based genomic DNA extraction kit from dried blood slices and a method for extracting genomic DNA from dried blood slices. Background technique [0002] With the development of various genetic testing technologies, dried blood film has gradually become one of the main samples for genetic testing, and the collection method of dried blood film has certain advantages over the traditional method of collecting whole blood. It requires less blood volume, can reduce the use of animals, facilitates blood sample collection, storage and transportation, and is an effective tool in the screening process of rare diseases. [0003] Genomic DNA contains all the genetic information in the cells. Extracting genomic DNA from dried blood film requires less blood volume, is convenient for collection and transportation, and has good DNA integrity. It is one of the...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/6806
CPCC12N15/1013C12Q1/6806
Inventor 林程忠陈荷萍吴涵杨呈勇
Owner GUANGDONG ASCENDAS GENOMICS TECH CO LTD
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