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Combination tag, combination tag joint, and application ofcombination tagand combination tag joint

A technology of tags and tag sequences, which is used in the construction of combined tag adapters, kits and gene libraries, and the field of combined tags, which can solve the problems that functional design cannot meet the needs of sequencing.

Pending Publication Date: 2019-10-15
TIANJIN MEDICAL LAB BGI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, these functional designs cannot meet the needs of sequencing, and further improvement is needed

Method used

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  • Combination tag, combination tag joint, and application ofcombination tagand combination tag joint
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  • Combination tag, combination tag joint, and application ofcombination tagand combination tag joint

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Combination label and its preparation

[0047] Example 1 provides a design solution for a combined label. Reference to the provided design plan figure 1 with 2 Shown:

[0048] 1) First, design the tag sequence according to the combined tag length of 6 and GC content of 50%;

[0049] 2) The tag sequence of length 6 is divided into three parts according to a group of 2 bases, each part has a GC content of 50%, so each part can get 8 AC, TG, CT, GA, AG, TC, CA, GT A short sequence, and the Hamming distance between two of the 8 sequences is greater than or equal to 1;

[0050] 3) Combining the sequences of the three parts, you can get 512 (8×8×8) labels with a length of 6, and the Hamming distance between the labels is at least 1, and the GC content of each label is still 50%. And there will not be three consecutive identical bases.

[0051] 4) Select and group 512 sequences. In the embodiment, each group contains 2 tags. Through grouping, the tags with the Hamming dista...

Embodiment 2

[0056] Example 2 Preparation method of combined label joint

[0057] Connect the combination tag designed in Example 1 and the adapter to obtain the combination tag adapter. Take the BGI-SEQ500 sequencing platform sequence as an example, order the following tag adapter sequence, where one end (3' end) of the combination tag adapter is designed as a single base Base overhanging the end, the single base overhanging end is T, which can be conveniently connected to the DNA with A at the end when building the library:

[0058] Table 1 Combination label connector

[0059]

[0060]

[0061] Take the combination label connector A as an example, refer to image 3 As shown, the combination label connector A contains two combination label connectors, namely A1 and A2. Take A1 as an example. image 3 The ACGTAC in the lower half of the box represents the tag sequence contained in the combinatorial tag linker, and the lower sequence in the same box represents the reverse complement of the tag seq...

Embodiment 3

[0065] Example 3 Evaluation of the improvement of sequencing depth by combination tag adapters and contamination between samples

[0066] 1. Obtain a DNA sample.

[0067] The test uses Horizon’s commercial standard products as DNA samples. The specific settings are shown in Table 2 below:

[0068] Table 2 Adapters used in different DNA samples

[0069] Test sample Standard product name Connector number Sample 1 HD777-1% mutation frequency YUC192_2*2AD_A Sample 2 HD777-1% mutation frequency YUC192_2*2AD_B Sample 3 HD777-1% mutation frequency YUC192_2*2AD_C Sample 4 HD786-1% mutation frequency YUC192_2*2AD_D Sample 5 HD786-1% mutation frequency YUC192_2*2AD_E Sample 6 HD786-1% mutation frequency YUC192_2*2AD_F Sample 7 HD777-negative YUC192_2*2AD_G Sample 8 HD777-negative YUC192_2*2AD_H Sample 9 HD777-1% mutation frequency Conventional connector-1 Sample 10 HD777-1% mutation frequency Conventional connector-2 Sample 11 HD777-1% mutation frequency Conventional connector-3 ...

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Abstract

The invention relates to the field of gene sequencing, in particular to a combination tag, a combination tagjoint, and application ofthe combination tagandthe combination tag joint. The combination tagcomprises two or more sets of tags, each set of the two or more sets of tags contains two or more tagsequences, the Hamming distance between any tag sequences in each of the two or more sets of tagsis greater than or equal to 2, the Hamming distance between any of the tag sequences between each of the two or more sets of tags is greater than or equal to 1. The combination tag joint contains a joint sequence and the combination tag. The combination tag or the combination tag jointis used for gene library construction, sequencing and the like, contamination between samples can be effectively avoided,and the sequencing depth is increased.

Description

Technical field [0001] The present invention relates to the field of gene sequencing, in particular to a combination tag, a combination tag adapter and applications thereof, in particular to a method for constructing a combination tag, a combination tag adapter, a kit and a gene library. Background technique [0002] Currently, DNA sequencing technology has been widely used in scientific research and medical fields. With the deepening of research and application, the sensitivity and specificity requirements are getting higher and higher, so some functional elements will be added to the DNA library adapter. Among them, there are two widely recognized designs: (1) UMI (Unique Molecular Identifier), which introduces a random sequence on the adapter, so that each DNA molecule to be detected carries a unique identification sequence, which can increase the sequencing depth; (2) ) DNA library dual index (Dual Index). A specific DNA sequence is attached to each end of each sample. Comp...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806C12N15/10C40B50/06C40B70/00C40B80/00
CPCC12Q1/6806C12N15/1093C40B50/06C40B70/00C40B80/00C12Q2525/191C12Q2531/113
Inventor 宋炎张娜刘玉朋邵林
Owner TIANJIN MEDICAL LAB BGI
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