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Bubble adapter elements and methods for constructing sequencing libraries using same

A technology of adapter elements and construction methods, applied in the biological field, can solve the problems of long overall library construction time, high requirements for initial library construction, and many PCR amplification times, and achieve simple connection methods, library construction cycle and cost reduction , the effect of high purification cost

Active Publication Date: 2021-08-06
MGI TECH CO LTD
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  • Summary
  • Abstract
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Problems solved by technology

Through the innovative bubbling joint element and its connection reaction technology in the solution and on the magnetic beads, the present invention focuses on solving the problems of too many joint connection steps and the number of PCR amplifications in the double joint library construction method of the CG sequencing platform. Many, the overall library construction time is long, the cost is high, and the initial quantity requirement for library construction is high

Method used

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  • Bubble adapter elements and methods for constructing sequencing libraries using same
  • Bubble adapter elements and methods for constructing sequencing libraries using same
  • Bubble adapter elements and methods for constructing sequencing libraries using same

Examples

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Embodiment 1

[0101] Example 1 Sequencing library construction of the present invention

[0102] 1. Genomic DNA interruption:

[0103] There are many ways to interrupt the genomic DNA of Yanhuang 1 cells, whether it is physical ultrasonic method or enzyme reaction method, there are very mature solutions on the market. This embodiment adopts the physical ultrasonic fragmentation method.

[0104] Take a 96-well PCR plate, add a polytetrafluoroethylene line, add 1ug of genomic DNA, add TE buffer solution or enzyme-free pure water to make up 100ul. After sealing the board, ultrasonically break it on the E220 ultrasonic breaker.

[0105] The interrupt conditions are set as follows:

[0106] Fill factor 21％ pressure (PIP) 500 Impulse factor 500 interrupt time 20s, 2 times

[0107] 2. Interrupt segment selection:

[0108] Magnetic bead purification or gel recovery can be used. The present embodiment adopts the magnetic bead purification method, specifically ...

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Abstract

The present invention provides a bubble adapter element and a method for constructing a sequencing library using the same. The linker element is a bubbling hybrid formed by a long nucleic acid chain A and a short nucleic acid chain B with complementary sequences at both ends and non-complementary intermediate sequences.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a bubble-shaped linker element, a method for constructing a sequencing library using the linker element, the constructed sequencing library and applications thereof. Background technique [0002] In the 1990s, since the launch of the capillary electrophoresis sequencer by AB Company and the launch of the Human Genome Project, DNA (Deoxyribonucleic acid, deoxyribonucleic acid) sequencing technology has undergone earth-shaking changes at an unimaginable speed. The second and third generation sequencers have also been launched one after another, facing the market. [0003] Among the second-generation sequencing platforms, the Blackbird sequencing platform of Complete Genomics (hereinafter referred to as CG) has higher sequencing accuracy (99.9998%) and greater sequencing throughput than other platforms. Applications in the field of research hold great promise. However, the lib...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/10C40B50/06C40B40/06C12Q1/6806C12Q1/6869
CPCC12Q1/6869C12Q2525/191C12Q2525/301C12Q2535/122C12Q1/68C12Q1/6855C12Q1/6874
Inventor 江媛赵霞李巧玲刘生茂王博陈利章文蔚蒋慧拉多杰·德马纳克
Owner MGI TECH CO LTD
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