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Library for bibasic sequencing quality assessment, reagent and application

A technology for next-generation sequencing and quality assessment, applied in libraries, chemical libraries, nucleotide libraries, etc., can solve unfavorable and troublesome problems, achieve the effects of reducing sequencing deviation, simple process, and high stability

Active Publication Date: 2018-06-26
MGI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method is cumbersome, and it is not conducive to targeted improvement of the error rate caused by each link in the sequencing process in the research and development of sequencing technology

Method used

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  • Library for bibasic sequencing quality assessment, reagent and application
  • Library for bibasic sequencing quality assessment, reagent and application
  • Library for bibasic sequencing quality assessment, reagent and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0049] In this example, a set of standard nucleic acid sequences containing base features such as high AT content, high GC content, poly structure, and hairpin structure are designed first, and then a library is designed for the standard nucleic acid sequence, and the BGISEQ linker sequence is added to the library sequence , index sequence and universal primer binding sequence. Artificially synthesize the designed library sequence, insert the artificially synthesized library sequence into pMD19-T plasmid, and introduce the plasmid into Ecoli Escherichia coli to make engineering bacteria. Extract plasmids from engineered bacteria to obtain library sequences for next-generation sequencing and evaluate sequencing quality. The details are as follows:

[0050] 1. Design of Standard Nucleic Acids

[0051] In this example, six standard nucleic acid sequences are designed according to the common recorded base characteristics in actual sequencing, such as high AT content, high GC con...

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PUM

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Abstract

The invention discloses a library for bibasic sequencing quality assessment, a reagent and an application. The library disclosed by the invention is a single-stranded DNA library of known sequences with different base characteristics, and adapter sequences and index sequences are connected into the library; and the single-stranded DNA library comprises at least one of high-AT-content single-stranded DNA, high-GC-content single-stranded DNA, poly-structured single-stranded DNA and hair-pin structured single-stranded DNA. According to the library disclosed by the invention, through sequencing byadopting the known sequences with the different base characteristics, the influence and deviation on sequencing caused by the different base characteristics can be assessed, the sequencing quality assessment can be achieved, the deviations can be corrected, and sequencing optimization is achieved. The invention provides a method for improving accuracy of nucleic acid sequencing. Through comparingsequencing results and the known sequences, sequencing deviations of different base characteristics are obtained, the improvement on a sequencing software algorithm is instructed, and the accuracy ofsequencing is improved; and the method provided by the invention can be used for effectively reducing the sequencing deviations and provides a simple and effective method for improving the accuracy of sequencing.

Description

technical field [0001] This application relates to the field of quality assessment of nucleic acid sequencing, in particular to a library, reagents and applications for quality assessment of next-generation sequencing. Background technique [0002] High-throughput sequencing technology is a very important technology that plays a vital role in both biological research and clinical applications, especially in precision medicine. As next-generation sequencing plays an increasingly important role in precision medicine, the corresponding requirements for its sequencing accuracy are also gradually increasing. The current mainstream next-generation sequencing platforms, such as illumina and proton, can achieve an accuracy rate of 99.9%, but the lengthening of sequencing reads and the complexity of base content may lead to a decline in sequencing accuracy. In order to better meet the role of next-generation sequencing in precision medicine, it is necessary to continuously improve s...

Claims

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Application Information

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IPC IPC(8): C40B40/06C12N15/70C12N1/21C12Q1/6869C12R1/19
CPCC12N15/70C12Q1/6869C40B40/06C12Q2535/122
Inventor 廖莎闫东东章文蔚徐崇钧陈奥陈莹赵杰许军强傅德丰何琳
Owner MGI TECH CO LTD