Abstract: A method for preparing an amplicon library for detecting the variation in a region to be tested of a target gene of a sample, including the following steps: 1) designing and synthesizing a forward outer primer F1, a forward inner primer F2, and a reverse primer R according to the target region; 2) carrying out a one-step PCR amplification on the sample to be tested using the forward outer primer F1, the forward inner primer F2, and the reverse inner primer R to obtain an amplified product, i.e., the amplicon library of the target region. This one-step library preparation technology can be applied to all second-generation platforms including IonTorrent, illumina and BGI/MGI platforms. Based on the library preparation method, the present invention has developed detection products for SNP, Ins/Del, CNV and methylation of DNA, as well as detection products for s gene fusion and expression of RNA samples.

Abstract: The present invention discloses a method for rapidly constructing amplicon library including the following steps: 1. Synthesizing a primer combination for constructing an amplicon library of a DNA sample, the primer combination of the amplicon library used to construct the DNA sample includes: a forward fusion primer designed according to the target amplicon, a reverse fusion primer designed according to the target amplicon, a forward universal primer and a reverse universal primer; 2. Constructing a PCR reaction system for the DNA sample; 3. Performing PCR. The method according to the present invention can be used to construct an amplicon library in a simple and rapid manner, and since a barcode is introduced before the start of PCR, the possibility of cross-contamination between the sample and the library is greatly reduced.

Abstract: The invention discloses a method for preparing an amplicon library for detecting a low-frequency mutation of a target gene. The invention provides a method for preparing an amplicon library for detecting a low-frequency mutation of a target gene, comprising the following steps: 1) design and synthesize a Barcode primer F1, an forward primer F2, a reverse outer primer R1, and a reverse inner primer R2; 2) perform a one-step PCR amplification on the cfDNA of the sample to be tested using the Barcode primer F1, the forward primer F2, the reverse outer primer R1, and the reverse inner primer R2 to obtain an amplified product, which is a DNA library for an amplicon sequencing. In addition to detect tissue samples, the method can also quickly, easily, sensitively and specifically amplify different target regions of cell free DNA from samples such as blood, urine, and CSF, and efficiently detect mutations as low as 0.1%.

Abstract: The present invention discloses a method for rapidly constructing amplicon library including the following steps: 1. Synthesizing a primer combination for constructing an amplicon library of a DNA sample, the primer combination of the amplicon library used to construct the DNA sample includes: an upstream fusion primer designed according to the target amplicon, a downstream fusion primer designed according to the target amplicon, an upstream universal primer and a downstream universal primer; 2. Constructing a PCR reaction system for the DNA sample; 3. Performing PCR. The method according to the present invention can be used to construct an amplicon library in a simple and rapid manner, and since a barcode is introduced before the start of PCR, the possibility of cross-contamination between the sample and the library is greatly reduced.