Method and kit for genotyping by utilizing high-throughput sequencing and an application of kit

A genotyping, high-throughput technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as high cost, high error rate, high error rate, etc., to achieve application Wide-ranging, fast, low-cost effects

Active Publication Date: 2020-08-18
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional approaches to genotyping using sequencing technologies often suffer from high data loss and error rates, especially at low read depths where detection from heterozygous loci and rare alleles yields high Error rate, and high cost

Method used

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  • Method and kit for genotyping by utilizing high-throughput sequencing and an application of kit
  • Method and kit for genotyping by utilizing high-throughput sequencing and an application of kit
  • Method and kit for genotyping by utilizing high-throughput sequencing and an application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0030] 1. Construct the universal Barcode fragment of the kit and amplify YFP DNA

[0031] (1) The Barcode fragment is used to identify different samples during data analysis, which is equivalent to the identification label of the sample. Specifically, it consists of an 18bp bridge fragment, an 8bp Barcode1 fragment, a 60bp sequence on YFP, an 8bp Barcode2 fragment, and a 20bp reverse fragment.

[0032] Taking DNA of 384 samples to be tested as an example, only 24 different Barcode1 sequences and 16 different Barcode2 sequences are needed to form 384 universal Barcode fragments for genotype identification of 384 samples.

[0033] The amplified universal Barcode fragment sequence designed in this example is shown in the following table, wherein, the first 18 bp of the front primer from the 5' end is a bridge fragment, the middle 8 bp is a Barcode1 fragment, and the last 20 bp is a sequence on YFP; the back primer The first 20 bp from the 5' end is a reverse fragment, the middl...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a method for genotyping by utilizing high-throughput sequencing, a kit and application of the kit. According to theinvention, a set of universal barcode fragments similar to the kit is constructed, so that genotype identification is carried out at a lower cost and a higher speed. According to the invention, a novel genotype identification technology developed by combining barocode and next-generation sequencing is quite efficient and simple for identification of SNP and InDel, the required cost is far lower than the market price, and especially the advantage of identification of large-batch samples is more outstanding. In addition, the technology can also be applied to gene localization, genetic map drawing and the like, so that the application range of the technology is quite wide.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method, kit and application for genotyping by high-throughput sequencing. Background technique [0002] High-throughput sequencing technology (High-throughput sequencing), also known as "next-generation" sequencing technology ("Next-generation" sequencing technology), can perform sequence determination and general reading on hundreds of thousands to several million DNA molecules in parallel at a time. Longer and shorter are symbols. High-throughput sequencing technology generally consists of four parts: sample preparation, library construction, sequencing reaction, and data analysis, which makes it possible to analyze the transcriptome and genome of a species in detail, so it is also called deep sequencing (deep sequencing). sequencing). Methods for SNP typing using high-throughput sequencing technology include: GBS (Genotyping-by-sequencing, genotyping technology bas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q2531/113C12Q2535/122
Inventor 李林李娟李明珠王席
Owner HUAZHONG AGRI UNIV
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