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

A high-throughput, labeling technology, applied in the field of nucleic acid labeling, can solve the problems of reducing the misallocation rate, affecting the experimental results, high misallocation rate, etc., and achieving the effect of low cost

Inactive Publication Date: 2011-07-06
XIAMEN UNIV
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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 b

Method used

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

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specific Embodiment

[0090] Specific examples are given below: Marking of 96 shellfish genome DNA samples

[0091] 1) Basic principles

[0092] 96-well plates in a 12×8 format according to Molecular Biology Standards (see figure 2 ), the specific tags can also be designed as a combination of 12×8. Since tags can be attached to both ends of the DNA, 96 DNA samples can be labeled by the specific combination of 12×8 tags at both ends. Name the label that marks different rows as column-barcode (c-bar for short), 8 in total, and the samples in the same row of the 96-well plate mark the same row label; name the label that marks different columns as column label (row -barcode), referred to as r-bar, 12 in total, the samples in the same column of the 96-well plate are marked with the same column label (see figure 2 ).

[0093] use T c and T r Indicate the length of the c-bar and r-bar labels respectively; use p D Indicates the probability of assigning sequence errors to other samples, specific to...

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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 ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 陈军柯才焕
Owner XIAMEN UNIV
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