Nucleic acid label for second-generation high-flux sequencing and design method thereof

Abstract

The invention discloses a nucleic acid label for second-generation high-flux sequencing and a design method thereof, and relates to a nucleic acid label. The invention provides the nucleic acid label for second-generation high-flux sequencing and the a design method thereof, which are capable of quickly, efficiently, specially and simultaneously labeling almost one hundred independent samples, applicable to the mixed sequencing of a plurality of samples and used under the condition with a 96-pore plate. The nucleic acid label comprises an A group including eight labels with the length of 5nt,a B group including eight labels with the length of 5nt and a C group including twelve labels with the length of 6nt. Designing a first tag sequence: Tag 1: CTAGA; designing other three tags: Tag 2: TGCAG; Tag 3: ACGTC; Tag 4: GATCT; designing other four tags: Tag 5: CGTAC; Tag 6: TAGCA; Tag 7: ATCGT; Tag 8: GCATG; and calculating a positional number with an identical basic group between every two tags.

Description

technical field [0001] The present invention relates to a nucleic acid tag, in particular to a nucleic acid tag for second-generation high-throughput sequencing and a design method thereof. Background technique [0002] Nucleic acid tag refers to a small piece of nucleic acid of known sequence, usually containing several to tens of nucleotides (nt), which is linked to the nucleic acid to be marked by molecular biological methods to mark the nucleic acid. The application of nucleic acid tags to multi-sample mixed sequencing can distinguish the sample from which each sequence comes. [0003] With the development and maturity of the second-generation DNA sequencing technology, the throughput of sequencing is getting higher and higher, which provides the possibility to measure multiple samples at the same time. For example, Roche / 454's GS FLX Titanium sequencing system can obtain 1 million sequences at a time. If 96 samples are measured at the same time, each sample can obtain ...

AI Technical Summary

Problems solved by technology

For 4nt tags, there are a total of 4^4=256 possible tags. If 96 of them are randomly selected as the actual tags, and the sequencing error rate is 0.8%, then a total of 3.2% tags will have sequencing errors , and each wrongly sequenced tag has a 37.5% (96 / 256=37.5%) possibility of being misread as other tags, that is, 1.2% of the total sequence will be wrongly assigned to other samples, such a high misassignment rate will Seriously affect the experimental results

Therefore, the label design must comply with scientific principles to reduce the misallocation rate

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