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Animal tissue DNA extracting kit based on paramagnetic particle method and application of animal tissue DNA extracting kit

A technology of animal tissues and kits, applied in the biological field, can solve the problems of long time consumption and cumbersome operation, and achieve the effects of low cost, simple extraction process, and increased extraction efficiency

Inactive Publication Date: 2016-03-23
NANJING INST OF ADVANCED LASER TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The DNA obtained by the above classical method is of high purity, which can meet the requirements of various experiments, but the operation is cumbersome and time-consuming, and a variety of instruments and equipment are required

Method used

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  • Animal tissue DNA extracting kit based on paramagnetic particle method and application of animal tissue DNA extracting kit
  • Animal tissue DNA extracting kit based on paramagnetic particle method and application of animal tissue DNA extracting kit
  • Animal tissue DNA extracting kit based on paramagnetic particle method and application of animal tissue DNA extracting kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1: Reagent preparation of animal tissue DNA extraction kit

[0047] (1) Preparation of lysate: first add a small amount of deionized water into the volumetric flask, add sodium lauryl sulfate with a concentration of 2% (m / v), add ethylenediaminetetraacetic acid with a concentration of 20mmol / L, Add tris at a concentration of 20mmol / L, add sodium chloride at a concentration of 1.4mol / L, add Triton X-100 at a concentration of 1% (v / v), and add deionized water to the Need volume, use sodium hydroxide and hydrochloric acid to adjust the pH value, make the pH value of the lysate 8.0, autoclave steam sterilization for 10 minutes;

[0048] (2) Preparation of binding solution: use anhydrous isopropanol;

[0049] (3) Preparation of washing solution Ⅰ: adding guanidine hydrochloride at a concentration of 10mol / L, adding ethylenediaminetetraacetic acid at a concentration of 40mmol / L, adding tris at a concentration of 40mmol / L, adding at a concentration of 2.8 The sodi...

Embodiment 2

[0054] Example 2: Using the kit in Example 1 to extract mouse liver DNA using method 1

[0055] Take 8 liver samples from the same mouse, each 50mg. The specific operation of the application is:

[0056] (1) Take 8 2mLEP tubes, add mouse liver to each tube, 300 μL of lysate, 10 μL of 20 mg / mL proteinase K, incubate in a water bath at 56°C for 30 minutes after homogenization, and then centrifuge;

[0057] (2) Take 8 new 2mLEP tubes, transfer the supernatant after centrifugation to the tube, add 20 μL of magnetic beads, 300 μL of binding solution, and combine for 10 minutes;

[0058] (3) Adsorb the magnetic beads to the tube wall with a magnet, and discard the liquid in the tube;

[0059] (4) Add 600 μL of washing solution I and mix well, use a magnet to adsorb the magnetic beads to the tube wall, and discard the liquid in the tube;

[0060] (5) Add 600 μL of washing solution II and mix well, use a magnet to adsorb the magnetic beads to the tube wall, and discard the liquid i...

Embodiment 3

[0068] Example 3: Use the kit in Example 1 to extract rat liver DNA using method 2

[0069] Take 8 liver samples from the same rat, each 50mg. The specific operation of the application is:

[0070] (1) Add 200 μL of binding solution to 1 row of 96-well reaction plate;

[0071] (2) Add 20 μL of magnetic beads and 600 μL of deionized water to row 2 of a 96-well reaction plate;

[0072] (3) Add 600 μL of washing solution I to the third row of the 96-well reaction plate;

[0073] (4) Add 600 μL of washing solution II to row 4 of the 96-well reaction plate;

[0074] (5) Add 600 μL of washing solution III to 5 rows of 96-well reaction plate;

[0075] (6) Add 100 μL of nucleic acid eluent to row 6 of a 96-well reaction plate;

[0076] (7) Take 8 2mLEP tubes, add 50 mg of rat liver, 300 μL of lysate and 10 μL of 20 mg / mL proteinase K to the tubes, and centrifuge directly after homogenization. Transfer the supernatant to row 1 of the 96-well reaction plate, run the nucleic acid e...

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Abstract

The invention discloses an animal tissue DNA extracting kit based on a paramagnetic particle method and application of the animal tissue DNA extracting kit. The kit comprises a lysis solution, a binding buffer, a cleaning solution I, a cleaning solution II, a cleaning solution III and a nucleic acid eluent. The lysis solution contains SDS. The three cleaning solutions all contain isopropanol. By combining the paramagnetic particle method with a traditional lysis solution for animal DNA extraction, broken cells are digested by protease K and SDS, pyrolysis nucleic acid is purified according to the principle of the paramagnetic particle method, and the kit is suitable for extracting animal tissue nucleic acid of various specifications in a laboratory, is also suitable for being commercially produced in batches, and has the advantages of being easy and rapid to operate, safe to use, high in extracting efficiency, low in manufacturing cost and the like; the extracting process of the kit can be manually completed and can also be automatically completed in a full-automatic nucleic acid extracting instrument, 1-16 animal tissue samples can be processed at a time through a 96-pore reaction plate during automatic extracting, and no other operation is needed in the extracting process.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a kit for extracting animal tissue DNA based on a magnetic bead method, which can be applied to DNA extraction from animal tissues of multiple species, and can also be applied to DNA extraction from different tissues of the same species. The invention also relates to a method for extracting DNA from animal tissue samples using the kit. Background technique [0002] Eukaryotic genomic DNA is widely used in the genetic breeding of animals and plants, the construction of gene maps, species identification, speciation and phylogenetic evolution. Whether it is genetic engineering or protein engineering, nucleic acid molecules are the main objects involved in the application of these technologies, so the separation and extraction of DNA is an important basic technology in molecular biology research. [0003] At present, there have been many studies and reports on the extraction of whole geno...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 孟庆海黄远福王炜
Owner NANJING INST OF ADVANCED LASER TECH
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