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Convertible adapters

An adaptor and nucleic acid molecule technology, which is applied in the field of generating asymmetrically labeled nucleic acid fragments and nucleic acid library templates, and can solve problems such as low efficiency and cumbersome processing steps.

Pending Publication Date: 2019-10-25
维尔道应用技术大学
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods include additional and tedious processing steps, or require long adapter oligonucleotides, which are less efficient in chemical synthesis and ligation than simple double-stranded adapters. Low

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  • Convertible adapters
  • Convertible adapters
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Examples

Experimental program
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Effect test

Embodiment

[0177] Transformable Adapter Design

[0178] Standard adapters for sequencing were chosen as templates to design transformable adapters according to the invention. Conventional IonTorrent adapters consist of two distinct double-stranded subunits that generate asymmetrically tailed nucleic acid pools:

[0179] adapter P1

[0180] 5'-CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT 3'

[0181]3’T*T*GGTGATGCGGAGGCGAAAGGAGAGATACCCGTCAGCCACTA-PO4 5’

[0182] * indicates phosphorothioate modification

[0183] Barcode Adapter A

[0184] 5'-CCATCTCATCCCTGCGTGTCTCCGACTCAGNNNNNNNNGAT 3'

[0185] 3’T*T*GGTAGAGTAGGGACGCACAGAGGCTGAGTCNNNNNNNNNCTA-PO4 5’

[0186] For better comparison with overhang-based ligation, T-tailed adapters were designed:

[0187] adapter P1-T

[0188] 5'-CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGATT 3'

[0189] 3’T*T*GGTGATGCGGAGGCGAAAGGAGAGATACCCGTCAGCCACTA-PO4 5’

[0190] *Indicates phosphorothioate modification and added T is not indicated

[0191] Barcode Adap...

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Abstract

The invention relates to a method and a kit comprising single or at least partially double-stranded adapters comprising sequences with base modifications that are ligated to nucleic acid fragments andconverted to generate asymmetric ends for specific recognition. The method is based on two main sequence conversion mechanisms, a direct conversion of specific bases and an indirect conversion by copying, wherein both mechanisms may be combined.

Description

[0001] The present invention belongs to the fields of biochemistry and diagnostics. It particularly belongs to the field of molecular biology and the generation of nucleic acid libraries, and more specifically to the generation of nucleic acid library templates for sequencing. Background technique [0002] Numerous methods in basic research, diagnostics, therapy, and forensics are based on nucleic acid analysis. More recently, highly parallel processes have been developed, such as microarrays and next-generation sequencing (also known as "massively parallel sequencing"), which allow simultaneous analysis of thousands, millions, or even more than billions of different nucleic acid molecules. For this, nucleic acids need to be converted into a general form to be processed in parallel as nucleic acid libraries. The optimal library format is one that provides common flanking sequences for unknown nucleic acids, so-called adapters. Since many analytical procedures require nucleic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12N15/10C12N15/11
CPCC12N15/1065C12Q1/6855C40B20/04C12Q1/6806C12Q2525/101C12Q2525/117C12Q2525/119C12Q2525/155C12Q2525/191C12Q2533/101
Inventor 耶恩·格洛克马库斯·弗罗姆
Owner 维尔道应用技术大学