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A universal adapter for multiple sequencing platforms, a library construction method and kit suitable for multiple sequencing platforms

A sequencing platform and universal adapter technology, applied in the field of high-throughput sequencing, can solve problems such as hindering in-depth detection research, waste of time, labor and material costs, etc.

Active Publication Date: 2022-03-18
SUZHOU BASECARE MEDICAL DEVICE CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When it is necessary to carry out experiments on multiple different sequencing platforms, it is necessary to construct multiple libraries suitable for different sequencing platforms for the same sample, such as figure 1 As shown, resulting in a waste of time and manpower and material resources
When the sample is scarce and cannot satisfy multiple database construction, it greatly hinders further in-depth detection research

Method used

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  • A universal adapter for multiple sequencing platforms, a library construction method and kit suitable for multiple sequencing platforms
  • A universal adapter for multiple sequencing platforms, a library construction method and kit suitable for multiple sequencing platforms
  • A universal adapter for multiple sequencing platforms, a library construction method and kit suitable for multiple sequencing platforms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Genomic DNA library construction and sequencing of known mutant cell lines

[0050] 1. Sample preparation

[0051] Select 9 cases of human lymphocyte line samples with known karyotype, including aneuploidy, samples with fragment deletions / duplications of different sizes (the minimum of which is about 1.5Mb), and normal control samples. Resuscitate, activate and expand the culture, and extract a sufficient amount of genomic DNA of the cell line as a test sample.

[0052] The DNA of 100ng is taken in every case sample and adopts the present invention figure 2 The indicated multi-sequencing platform universal adapters were used for library construction. Take 100ng of DNA from each sample and complete it with the standard library construction method (traditional library construction method) published on the official website of Thermo Fisher, as a control. For specific method steps, refer to the official website of Thermo Fisher (https: / / www.thermofisher.com) ....

Embodiment 2

[0069] Example 2 Library construction and sequencing of single-cell samples of known variant cell lines

[0070] 1. Sample preparation

[0071] Select 10 samples of human lymphoid cell lines with known karyotypes, including aneuploidy, samples with fragment deletions / duplications of different sizes (the minimum of which is about 1.5Mb), and normal control samples. When it is resuscitated, activated and cultured to the best state, a single cell is picked as a single cell detection sample. Single-cell samples were first subjected to whole-genome amplification using the PicoPlex method, and then the amplified products were used for high-throughput library construction and sequencing. For specific PicoPlex method steps, please refer to the official website of TaKaRa Company (https: / / www.takarabio.com / ).

[0072] 2. Rapid fragmentation / end repair / dA tail addition of genomic DNA

[0073] Put each reagent on ice, prepare the reaction system in a new PCR tube according to the table...

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Abstract

The invention relates to a multi-sequencing platform universal adapter, a library construction method and a kit suitable for the multi-sequencing platform, based on the design of the multi-sequencing platform universal adapter and the development of a supporting rapid library construction process, DNA fragmentation, terminal Repair and dA tail addition, and then connect the two ends of the DNA fragments with multi-sequencing platform universal adapters, and the constructed library can be simultaneously sequenced on multiple sequencing platforms for high-throughput sequencing. This method solves the clinical problem of multiple construction of different sequencing platform libraries due to differences in sequencing platforms, and greatly reduces the time and cost of library construction. At the same time, the low initial template amount required for a single library construction also greatly saves the consumption of clinical samples .

Description

technical field [0001] The invention relates to the field of high-throughput sequencing, in particular to a multi-sequencing platform universal adapter, a library construction method and a kit suitable for the multi-sequencing platform. Background technique [0002] With the development of high-throughput sequencing technology, research in various bioscience fields and testing in clinical institutions involve the construction of sequencing libraries and high-throughput sequencing services. The promotion of clinical applications has driven the development of the high-throughput sequencing market, and has also created higher requirements for library construction and sequencing efficiency. The time and cost of library preparation have become the key factors for the widespread promotion of high-throughput sequencing applications. The preparation of high-throughput DNA library samples is the process of connecting the two ends of the DNA fragments to be sequenced with adapter sequ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6806C40B50/06C40B80/00
CPCC12Q1/6806C40B50/06C40B80/00C12Q2525/191C12Q2531/113C12Q2535/122
Inventor 赵丁丁孔令印冒燕夏琴梁波
Owner SUZHOU BASECARE MEDICAL DEVICE CO LTD
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