Universal adapters of multiple sequencing platforms, library construction method suitable for multiple sequencing platforms and kit

A sequencing platform and a technology of universal joints, applied in the field of high-throughput sequencing, can solve the problems of wasting time, manpower and material resources, hindering in-depth detection and research, etc.

Active Publication Date: 2020-05-08
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 f

Method used

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  • Universal adapters of multiple sequencing platforms, library construction method suitable for multiple sequencing platforms and kit
  • Universal adapters of multiple sequencing platforms, library construction method suitable for multiple sequencing platforms and kit
  • Universal adapters of multiple sequencing platforms, library construction method suitable for multiple sequencing platforms and kit

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 universal adapters of multiple sequencing platforms, a library construction method suitable for the multiple sequencing platforms and a kit. Based on design of the universal adapters of the multiple sequencing platforms and development of a matched fast library construction process, DNA fragmentation, end repair and dA tailing adding are completed through a one-step reaction, and then two ends of a DNA fragment are connected with the universal adapters of the multiple sequencing platforms so that a constructed library can conduct high-throughput sequencing on the multiple sequencing platforms at the same time. By means of the method, the clinical problem that due to difference of sequencing platforms, different sequencing platform libraries need to be constructed for multiple times is solved, library duration is greatly shortened, the library construction cost is greatly reduced, and meanwhile, due to a low demand for the starting template amount of single library construction, consumption of clinical samples is greatly reduced.

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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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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