DNA library construction method for fetus chromosome aneuploid detection

A technology for aneuploidy and library construction, which is applied in the field of non-invasive prenatal screening and detection, can solve the problems of high cost of library construction methods, poor practical operation results, and difficult replication operations, etc., so as to reduce the sequencing process and improve labor efficiency , time-saving effect

Inactive Publication Date: 2018-10-12
CHANGSHA KINGMED MEDICAL DIAGNOSTICS INST
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Problems solved by technology

[0008] The main problem solved by this application is to provide a DNA library construction method in the detection of fetal chromosomal aneuploidy, which has simple operation, scientific process, clear standards, and good practical operation effect, so as to solve the The DNA library construction method has high cost, unscientific process, unclear standards, poor practical operation effect, high operation difficulty, and technical problems that are difficult to replicate

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  • DNA library construction method for fetus chromosome aneuploid detection

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Embodiment 1

[0039] A DNA library construction method in the detection of fetal chromosomal aneuploidy, characterized in that, comprising:

[0040] Step 1: Physically interrupt the sample DNA to obtain a fragmented DNA solution;

[0041] Step 2: performing end repair and magnetic bead purification on the fragmented DNA to obtain purified blunt-ended DNA;

[0042] Step 3: performing adapter ligation on the blunt-ended DNA to obtain adapter-ligated DNA;

[0043] Step 4: performing PCR amplification on the DNA connected with the adapter to obtain a library;

[0044]Wherein, the step three specifically includes: sequentially adding blunt-end DNA, nuclease-free water, ligation buffer, dNTP, DNA ligase, P1 linker and tag sequence X into the centrifuge tube and mixing, and centrifuging and purifying after reaction.

[0045] The third step specifically includes:

[0046] Step 1: Add 25ul blunt-end DNA, 16ul nuclease-free water, 5ul ligation buffer, 1ul dNTP, 1ul DNA ligase, 1ul P1 linker and 1u...

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Abstract

The invention discloses a DNA library construction method for fetus chromosome aneuploid detection. The method comprises the steps of 1, performing physical interrupting on a sample DNA, and obtaininga fragmented DNA solution; 2, performing tail end repairing and magnetic bead purification on the fragmented DNA, and obtaining the purified blunt end DNA; 3, performing joint connection on the bluntend DNA, and obtaining DNA of the joint; 4, performing PCR amplification on the DNA of the joint, and obtaining a library, wherein according to step 3, the blunt end DNA, nuclear-free water, connection buffer solution, dNTP, DNA ligase, P1 joint and label sequences X are sequentially added into the centrifugal tube and mixed to be uniform, and centrifuging and purifying are performed after the reaction; operation is easy, the flow path is scientific, the standard is clear, and the practical operation effect is good.

Description

technical field [0001] The invention belongs to the field of non-invasive prenatal screening and detection, and specifically refers to a DNA library construction method in the detection of fetal chromosome aneuploidy. Background technique [0002] The rapid acquisition of genetic information of living organisms is of great significance to the research of life sciences. The reliance of the first-generation sequencer on electrophoretic separation technology makes it difficult to further improve the analysis speed and parallelization degree, and it is also difficult to reduce the sequencing cost through miniaturization. After continuous technical development and improvement, at the beginning of the 21st century, the second-generation sequencing technology marked by Roche454 technology, Illumina's GenomeAnalyzer technology and ABI's Solid technology was born. Second-generation technology has greatly reduced the cost of sequencing, and has also greatly increased the speed of seq...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C40B50/06
CPCC12Q1/6806C40B50/06C12Q2525/191C12Q2563/143C12Q2563/149
Inventor 何媛周梅华
Owner CHANGSHA KINGMED MEDICAL DIAGNOSTICS INST
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