Kit for extracting blood genome DNA based on magnetic bead method and use method for kit

A technology of genome and magnetic bead method, which is applied in the field of molecular biology, can solve the problems of increased workload of experimenters and increased extraction time, and achieve the effects of busy procedures, improved work efficiency, and high DNA extraction yield

Inactive Publication Date: 2016-04-27
SUZHOU ENRICHING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method can quickly extract a large volume of blood samples, as the number of samples increases, the extraction time will increase significantly due to the pretreatment steps of red blood cells, and the workload of the experimenters will also increase accordingly.

Method used

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  • Kit for extracting blood genome DNA based on magnetic bead method and use method for kit
  • Kit for extracting blood genome DNA based on magnetic bead method and use method for kit
  • Kit for extracting blood genome DNA based on magnetic bead method and use method for kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A kit for extracting blood genomic DNA based on the magnetic bead method, comprising independently packaged:

[0035] Cell lysate: a mixed solution of 6M guanidine hydrochloride, 0.5M sodium chloride and 1% Tween 20 by volume fraction. Specific preparation method: Weigh 573g of guanidine hydrochloride and 29g of sodium chloride, add 800mL of deionized water and stir to dissolve, add 10mL of Tween 20, continue to add deionized water to make up to 1L.

[0036] Proteinase K solution: Proteinase K solution is a mature commercial product, which is a serine protease with broad cleavage activity, purified from Candida albicans Limberella. The proteinase K solution is a solution obtained by dissolving proteinase K in water.

[0037] Magnetic bead dispersion: magnetic bead suspension with a concentration of 20mg / mL, the core of the magnetic bead is ferric oxide (Fe 3 o 4 ), the surface is coated with SiO 2 layer. Specific preparation method: Weigh 2g of magnetic beads, add ...

Embodiment 2

[0055] Use the kit of Example 1 to carry out high-throughput extraction experiments on 16 parts of blood genomic DNA:

[0056] Step 1: Thaw the frozen blood sample at 4°C or at room temperature; stabilize the water bath to 58°C; set the vibration frequency of the vortex mixer to 1150rpm.

[0057] Step 2: Add 200 μL of blood sample, 10 μL of proteinase K solution, and 200 μL of cell lysate to the 48-well plate in sequence with a multi-channel automatic pipette, 16 samples in parallel, vortex for 30 seconds to mix well, and place in a water bath Lyse for 10 minutes.

[0058] Step 3: Take out the 48-well plate, add 100 μL of magnetic bead dispersion and 300 μL of isopropanol to the 16 sample wells with a multi-channel automatic pipette, place the 48-well plate in a vortex mixer for 5 minutes, and place Place the 48-well plate on the 48-well plate magnetic plate rack and let it stand for 20 seconds until the magnetic beads are completely adsorbed. Keep the 48-well plate fixed on ...

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Abstract

The invention relates to a kit for extracting blood genome DNA based on a magnetic bead method. The kit comprises a cell lysis solution, a proteinase K solution, a magnetic bead dispersion solution, a first cleaning solution, a second cleaning solution and an elution solution, wherein the cell lysis solution comprises a guanidine compound, sodium chloride and a tween 20; a magnetic bead in the magnetic bead dispersion solution has the characteristics that a core is superparamagnetic ferroferric oxide, the surface is coated with silicon hydroxyl, and the particle size is 200-800nm; the first cleaning solution comprises Tris alkali, EDTA, isopropanol and water; the second cleaning solution is an ethanol solution; and the elution solution is Tris alkali or de-ionized water. The kit is simple to operate and can greatly improve working efficiency of blood genome DNA extraction; without pretreating erythrocytes, the cell lysis solution is directly adopted for pyrolyzing blood cells, and the magnetic bead dispersion solution can be added and combined with the genome DNA; and the kit is high in extraction volume and suitable for high-throughput experiments.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a kit for extracting blood genome DNA based on a magnetic bead method and a use method thereof. Background technique [0002] In recent years, with the rapid development of molecular biology technology, the research at the genome level has become a hot spot for researchers. Many routine experiments in molecular biology need to extract genomic DNA as a prerequisite, whether it is gene sequencing, PCR amplification, or construction of BCA library, etc., and the concentration, purity, molecular weight range and integrity of the primary structure of the extracted genomic DNA Will affect downstream molecular biology experiments. In recent years, precision medicine and in vitro diagnostic technology have developed vigorously, and the amount of samples to be processed has become increasingly large. Therefore, efficient, convenient, reliable, and high-throughput DNA extracti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 吴巧胡馨予赵青玲尚春庆曲峰
Owner SUZHOU ENRICHING BIOTECH CO LTD
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