Method for correcting sequence abundance deviation of secondary high-flux sequence test by DNA sequence bar codes

Abstract

The invention relates to a method for correcting the sequence abundance deviation of secondary high-flux sequence test by DNA sequence bar codes. Through introducing ten or more than ten DNA sequences to be used as bar code labels, the DNA of various different samples is corrected through abundance data obtained through the high-flux sequence test. The invention introduces the DNA sequence bar codes through three different designs, and the three methods can be applicable to various different sample conditions. During the three technical designs are described, the design uses 454 high-flux sequence test as the example. However, the principle is also applicable to other high-flux sequence test technical platforms, including SOLiD, Polynator and the like. The invention effectively overcomes the experimental error introduced by the emulsion PCR steps in building databases of several platforms of the current secondary high-flux sequence test. The technical method and the experimental flow process designed by the invention distinguish and correct the higher sequence read-out abundance of some template sequence in the emulsion PCR process caused by crack pollution of water-in-oil small balls through introducing more than one million kinds of (410=1048576) DNA sequence bar codes.

Description

Technical field: [0001] The invention relates to a method for correcting the sequence abundance deviation of the second-generation high-throughput sequencing by using a DNA sequence barcode. Background technique: [0002] High-throughput sequencing technology is a revolutionary change to traditional sequencing. It can sequence hundreds of thousands to millions of DNA molecules at a time, making it possible to analyze the transcriptome and genome of a species in detail. [0003] The development of next-generation high-throughput sequencing technology started with Genome Sequencer20 System (1), an innovative ultra-high-throughput genome sequencing system based on pyrosequencing and emulsion PCR, launched by 454 at the end of 2005. In 2007, 454 launched a second-generation genome sequencing system with better performance: Genome Sequencer FLX System. At present, besides the 454 GS system, the second-generation high-throughput sequencing technology platforms that have been comm...

AI Technical Summary

Problems solved by technology

A major defect of the emulsion PCR technology is that the water-in-oil pellets inevitably rupture during the PCR reaction and contaminate other blank pellets, which leads to falsely increas

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