Kit for extracting nucleic acid from bacteria by using paramagnetic particle method and extracting method

An extraction method, the technology of the magnetic bead method, is applied in the field of kits for extracting nucleic acid from bacteria based on the magnetic bead method, which can solve the problems of time-consuming and low efficiency, and achieve the effects of reducing cross-contamination, easy operation, and reducing physical damage

Inactive Publication Date: 2015-03-25
NANJING ZHONGKE SHENGUANG TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Extraction techniques such as alkaline lysis extraction and column purification are currently widely used in the extraction of bacterial DNA and RNA, but these methods still have many disadvantages, such as long time-consuming, low efficiency, and due to the use of toxic organic solvents such as phenol, the operation is difficult. certain risk

Method used

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  • Kit for extracting nucleic acid from bacteria by using paramagnetic particle method and extracting method
  • Kit for extracting nucleic acid from bacteria by using paramagnetic particle method and extracting method

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Effect test

Embodiment 1

[0033] 1. The preparation method of nucleic acid extraction kit and its reagents in bacteria:

[0034] ①Preparation of bacterial lysate: first add a small amount of sterile water into the volumetric flask, add sodium lauryl sulfate with a concentration of 55mmol / L, add ethylenediaminetetraacetic acid with a concentration of 25mmol / L, and add a concentration of 100mmol / L L of trishydroxymethylaminomethane, adding sodium chloride with a concentration of 500mmol / L, adding sterile water to the required volume, using sodium hydroxide to adjust the pH value so that the pH value of the bacterial lysate is 8.0, high-pressure steam Sterilize for 10 minutes.

[0035] ②Preparation of magnetic bead binding solution: first add a small amount of sterile water into the volumetric flask, add polyethylene glycol-8000 with a concentration of 25mmol / L, and sodium chloride with a concentration of 3mol / L, and then add sterile water to the desired concentration. volume, autoclaved for 10 minutes. ...

Embodiment 2

[0046] 1. The preparation method of nucleic acid extraction kit and reagents thereof in bacteria: with embodiment 1.

[0047] 2. Application of the kit:

[0048] Bacterial culture is with embodiment 1;

[0049] Take 1ml of Escherichia coli bacteria solution and add it to the EP tube, centrifuge at 13000r / min for 2 minutes, discard the supernatant, add 300μL of bacterial lysis solution, and mix well; add the above liquid to row 1 and row 7 of a 96 deep-well reaction plate , and add 300 μL of ethanol; add 100 μL of magnetic beads, 100 μL of magnetic bead binding solution, and 400 μL of double distilled water to row 2 and row 8 of a 96 deep-well reaction plate; add 600 μL of magnetic bead washing solution to row 3 of a 96 deep-well reaction plate - Row 5, row 9-11; add 100 μL of nucleic acid eluent to row 6 and row 12 of the 96 deep-well reaction plate; put the 96 deep-well reaction plate into the nucleic acid extractor, set the lysis time to 5 minutes, and the binding time to 5...

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Abstract

The invention discloses a kit for extracting nucleic acid from bacteria by using a paramagnetic particle method and an extracting method. The kit comprises a bacteria lysate, a magnetic bead binding solution, a magnetic bead scrubbing solution and a nucleic acid eluent. The bacteria lysate comprises sodium dodecyl sulfate, ethylenediaminetetraacetic acid, tri-hydroxymethyl aminomethane and sodium chloride; the magnetic bead binding solution comprises polyethylene glycol-8000 and sodium chloride; the magnetic bead scrubbing solution comprises ethanol; and the nucleic acid eluent comprises tri-hydroxymethyl aminomethane and ethylenediamine tetraacetic acid. The kit comprises the unique bacteria lysate which has a strong lysis function for the bacteria; and the kit can be used for manual extraction and instrument extraction by using a commercially available nucleic acid isolation machine, high-purity and high-concentration bacteria nucleic acid can be extracted.

Description

technical field [0001] The invention relates to the biotechnology field of nucleic acid extraction, in particular to a kit for extracting nucleic acid from bacteria based on a magnetic bead method. The invention also relates to a method for extracting nucleic acid from bacteria using the magnetic bead method. Background technique [0002] Polymerase Chain Reaction (PCR) technology has been more and more used in the field of molecular biology since its appearance in the 1980s. The molecular biology technology of nucleic acid is used to detect pathogenic microorganisms, construct gene banks, origin of species, It is one of the commonly used research methods for diversity assessment, phylogenetic relationship and phylogenetic evolution. However, the extraction of nucleic acid is the prerequisite for the application of PCR technology, and the ability to extract high-quality nucleic acid molecules is the key to many nucleic acid molecular biology experiments. The sensitivity and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 王炜孟庆海李智洋何农跃曲文静
Owner NANJING ZHONGKE SHENGUANG TECH
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