Method for constructing dna library that can be used for non-invasive prenatal detection of single gene disease
A DNA library, single-gene disease technology, applied in libraries, chemical libraries, nucleotide libraries, etc., can solve the problems of clearing and cyclization of single molecules, which are not easy to accurately quantify, and the molecules cannot be cyclized, and improve the efficiency of cyclization. , The effect of cyclization effect is easy to monitor
Active Publication Date: 2021-07-30
ZHEJIANG ANNOROAD BIO TECH CO LTD +2
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Problems solved by technology
However, in the prior art, the denatured ssDNA has the possibility of renaturation during the circularization process, resulting in a considerable portion of molecules that cannot be circularized and are eliminated by subsequent linear digestion
In addition, it is not easy to accurately quantify circularized single molecules, resulting in the success of circularization can only be evaluated after sequencing, which may cause a lot of waste
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[0058] The present invention will be described more specifically by way of examples below. It should be understood that the embodiments described here are used to explain the present invention, not to limit the present invention.
[0059] sample selection
[0060] First, press figure 1 SNP (rs1801394) related to folic acid metabolism in fetal DNA and maternal blood DNA was sequenced by generation sequencing. The plasma of two samples was selected for non-invasive experiments. The genotypes of the mothers corresponding to the two samples were both AG, and the genotypes of the fetuses were AA (sample 1) and GG (sample 2), respectively.
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The invention provides a method for constructing a DNA library that can be used for non-invasive prenatal monogenic disease detection. In the method for constructing a DNA library for sequencing of the present invention, the step of performing PCR amplification on the adapter-added DNA fragments to obtain the first amplification product is performed, and the first amplification product is digested with the restriction endonuclease , the step of obtaining enzyme-cleaved products, and the step of self-ligating the enzyme-cleaved products to obtain a DNA mixture containing double-stranded circularized products.
Description
technical field [0001] The invention belongs to the field of molecular diagnosis, and in particular relates to a method for constructing a DNA library that can be used for non-invasive prenatal monogenic disease detection. Background technique [0002] The presence of fetal free DNA (cell-free fetal DNA, cffDNA) in the plasma of pregnant women makes it possible to detect the genetic information of the fetus by non-invasive means. In view of this, more and more methods are used to non-invasively detect the genetic disease status of the fetus. Among them, the use of next-generation sequencing to screen for abnormal chromosome numbers has been recognized and industrialized by the state, including trisomy of chromosomes 13, 18, and 21. Recently, with the development of technology, non-invasive methods have been used to detect chromosome microdeletions and microduplications. However, the non-invasive detection of single-gene disease still needs to be further developed technical...
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IPC IPC(8): C12Q1/6883C40B40/08C40B50/06
CPCC12Q1/6806C40B40/08C40B50/06
Inventor 惠峰洪燕杜洋彭胜斌玄兆伶李大为梁峻彬陈重建
Owner ZHEJIANG ANNOROAD BIO TECH CO LTD
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