Method for constructing dna library that can be used for non-invasive prenatal detection of single gene disease

Abstract

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...

AI Technical Summary

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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