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Binding solution for purification of nucleic acid and application of binding solution

A nucleic acid and buffer technology, applied in the field of purifying nucleic acid binding solution, can solve problems to be improved, etc.

Inactive Publication Date: 2017-11-03
TIANGEN BIOTECH BEIJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the existing methods for separating and purifying nucleic acids still need to be improved

Method used

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  • Binding solution for purification of nucleic acid and application of binding solution
  • Binding solution for purification of nucleic acid and application of binding solution
  • Binding solution for purification of nucleic acid and application of binding solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A 200bp non-specifically amplified fragment was removed from a 2kb polymerase chain reaction (PCR) product. The specific operation steps are as follows:

[0040] (1) Add 0.6 times the volume of binding solution (the binding solution contains 2.5mol / L potassium chloride, 20V% tetraethylene glycol and 100mMTRis, and the pH value is 7.5) and 5 μl of 1 μm average particle size to 40 μl of the sample to be recovered and Surface-modified magnetic beads with carboxyl functional groups, mixed thoroughly, and oscillated for 5 minutes to facilitate the binding of the magnetic beads to the target fragment;

[0041] (2) Place the centrifuge tube in step (1) on the magnetic stand for 2 minutes, and carefully remove the liquid when the magnetic beads are completely absorbed;

[0042] (3) Place the centrifuge tube in step (2) on the magnetic stand, add 300 μl of rinsing solution (the rinsing solution contains 75% ethanol), and do not disturb the magnetic beads;

[0043] (4) Place th...

Embodiment 2

[0050] The 100 bp non-specific amplification band was removed from the 300 bp polymerase chain reaction (PCR) product. The specific operation steps are as follows:

[0051] (1) Add 1 volume of binding solution (the binding solution contains 3.5mol / L sodium chloride, 30V% polyethylene glycol 4000 and 100mM3-MOPS, pH value is 7.0) and 5μl average particle size to 50μl of the sample to be recovered. Magnetic beads with a diameter of 1 μm and surface-modified carboxyl functional groups were mixed thoroughly and shaken for 5 minutes to facilitate the binding of the magnetic beads to the target fragment;

[0052] (2) Place the centrifuge tube in step (1) on the magnetic stand for 2 minutes, and carefully remove the liquid when the magnetic beads are completely absorbed;

[0053] (3) Place the centrifuge tube in step (2) on a magnetic stand, add 300 μl of rinsing solution (the rinsing solution contains 70% ethanol), and do not disturb the magnetic beads;

[0054] (4) Place the cent...

Embodiment 3

[0061] Purify and recover the small amount of circulating nucleic acid after the linker is added during the construction of the next-generation sequencing library. The specific operation steps are as follows:

[0062] (1) Add 50 μl of binding solution (the binding solution contains 3.5M potassium chloride, 25 mass % PEG6000 and 100 mMT Ris, pH value is 9) and 5 μl of the product with an average particle size of 1 μm and surface modification For magnetic beads with carboxyl functional groups, shake for 5 minutes to facilitate the full binding of the magnetic beads to the target fragment;

[0063] (2) Place the centrifuge tube in step (1) on the magnetic stand for 2 minutes, and carefully remove the liquid when the magnetic beads are completely absorbed;

[0064] (3) Place the centrifuge tube in step (2) on a magnetic stand, add 300 μl of rinsing solution (the rinsing solution contains 80% ethanol), and do not disturb the magnetic beads;

[0065] (4) Place the centrifuge tube ...

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Abstract

The invention relates to a binding solution for purification of nucleic acid. The binding solution particularly comprises salts and solubilizers; the salts are selected from at least one of potassium chloride, potassium acetate, lithium chloride, lithium acetate, sodium chloride and sodium acetate; the solubilizers are nonionic solubilizers and / or ionic solubilizers. A method of separating and purifying nucleic acid via the binding solution is simple to perform and has no need for centrifuging, the target nucleic acid purified has good purity and is directly applicable to linking, transformation, enzyme cleavage, sequencing, hybridizing and other molecular biological experiments.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular, the invention relates to a binding liquid for purifying nucleic acid and its application. Background technique [0002] The classic nucleic acid separation and purification method is to adsorb target nucleic acid molecules on the silicon matrix membrane under the action of high-concentration chaotropic salts, while non-target nucleic acids, proteins, salts and organic substances are not adsorbed, so as to achieve the purpose of separation and purification. This method can be divided into two categories according to the purpose of purification: direct purification method and gel cutting recovery method. The direct purification method is suitable for the case where the product is a single DNA fragment or all DNA fragments in the solution need to be recovered; and the gel cutting recovery method Suitable for selective recovery of DNA in solution. From the aspect of the molecular characte...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 崔黎刘玉方李晓晨孙克非
Owner TIANGEN BIOTECH BEIJING
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