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Splitting decomposition composition, application thereof, kit, method for preparing nucleic acid through splitting decomposition composition and method for analyzing nucleic acid

一种核酸分析、组合物的技术，应用在生物化学设备和方法、测试用样品的制备、DNA制备等方向，能够解决样品交叉污染等问题，达到耗时短、流程简单、减少样品交叉污染的效果

Inactive Publication Date: 2016-04-06
HEALTH&HELP BIOSCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, although the method of separating DNA by magnetic beads requires fewer centrifuge tubes, it has multiple steps and requires expensive special equipment.
These existing techniques require multiple replacement of centrifuge tubes or multiple washes to collect DNA, which easily leads to sample cross-contamination
There are very limited reports of methods that directly perform real-time PCR without extracting DNA.

Method used

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  • Splitting decomposition composition, application thereof, kit, method for preparing nucleic acid through splitting decomposition composition and method for analyzing nucleic acid
  • Splitting decomposition composition, application thereof, kit, method for preparing nucleic acid through splitting decomposition composition and method for analyzing nucleic acid
  • Splitting decomposition composition, application thereof, kit, method for preparing nucleic acid through splitting decomposition composition and method for analyzing nucleic acid

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Effect test

Embodiment 1

[0081] The preparation of embodiment 1 cracking composition

[0082] Component A: NaOH with a concentration of 200mmol / L;

[0083] Component B: PEG with a concentration of 1 mg / mL (molecular weight is 2000);

[0084] The molar ratio of A and B is: 150:0.75;

[0085] A lysis composition is prepared.

Embodiment 2

[0086] The preparation of embodiment 2 cracking composition

[0087] Component A: Ca(OH) with a concentration of 0.1mmol / L 2 ;

[0088] Component B: PEG with a concentration of 200 mg / mL (molecular weight: 100);

[0089] The molar ratio of A to B is 0.1:1.05;

[0090] A lysis composition is prepared.

Embodiment 3

[0091] The preparation of embodiment 3 cracking composition

[0092] Component A: KOH with a concentration of 100mmol / L;

[0093] Component B: PEG with a concentration of 100 mg / mL (molecular weight is 1000);

[0094] The molar ratio of A and B is: 200:0.45;

[0095] A lysis composition is prepared.

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Abstract

The invention relates to the technical field of biology, in particular to a splitting decomposition composition, an application thereof, a kit, a method for preparing nucleic acid through the splitting decomposition composition and a method for analyzing the nucleic acid. An obtained splitting decomposition product can be directly used for the method for analyzing the nucleic acid, and the nucleic acid does not need to be purified. By means of the splitting decomposition composition, the nucleic acid in cells is discharged into a solution in the mode that splitting decomposition is carried out on multiple kinds of biological tissues and the multiple cells; after the reaction is ended, the nucleic acid is not subjected to the step of conventional nucleic acid separation and purification, and the nucleic acid in the whole-cell splitting decomposition mixed state is directly used as a template to guide nucleic-acid amplification and analyzing such as nucleic-acid amplification carried out through real-time fluorescence qPCR.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a lysing composition and its use, a kit, a method for preparing nucleic acid by using the lysing composition, and a method for analyzing nucleic acid. Background technique [0002] Fluorescent quantitative PCR (qPCR) is a DNA amplification technology based on polymerase chain reaction PCR (Polymerase Chain Reaction) which has been improved in many aspects. Quantitative PCR is performed in a thermalcycler instrument by emitting a beam of light of a specific wavelength to illuminate each DNA sample and detecting the fluorescence emitted by an excited fluorophore (fluorophonre). qPCR requires the use of fluorescent probes or fluorescent primers, and the amount of target DNA changes during real-time amplification can be monitored simultaneously by a real-time PCR instrument. In addition, qPCR can also amplify one or more target DNA sequences simultaneously. [0003] qPCR (quantitativeP...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/68
CPCC12N15/1003C12Q1/6806C12N15/10G01N1/28
Inventor 王晓惠孙冬捷庄青青
Owner HEALTH&HELP BIOSCI
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