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Biochemical micro-reaction system, high-throughput sequencing database builder and application

A micro-reaction system and biochemical technology, applied in the field of molecular biology, can solve the problems of consuming disposable consumables and high reagent costs.

Active Publication Date: 2016-08-03
李星军
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the existing library construction methods, in order to ensure a relatively high concentration of target samples, the usual sample volume range is 200 μL, and the reagent volume is greater than 200 μL. The relatively large reaction volume will lead to high reagent costs, and it also requires a large amount of one-time use. Consumables (such as gun tips, etc.)

Method used

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  • Biochemical micro-reaction system, high-throughput sequencing database builder and application
  • Biochemical micro-reaction system, high-throughput sequencing database builder and application
  • Biochemical micro-reaction system, high-throughput sequencing database builder and application

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application example 1

[0135] Application Case 1: High-throughput library preparation of common DNA samples

[0136] use figure 1 The library construction instrument shown uses the plasma cfDNA sample of a 12-week pregnant woman as a gene sample, and uses the IonPlusFragmentLibraryKit kit of Life Company to prepare a high-throughput sequencing library. The specific operation process is as follows:

[0137] (1) End repair, establishment of biochemical micro-reaction system 1:

[0138] The constant flow pump 5 is used to sequentially draw the first oily liquid and the second oily liquid into the reaction hole position 61 of the reaction well plate 6, and the reaction well plate 6 is a temperature control cavity of the PCR instrument, wherein the first oily liquid ( Heavy oil) is specifically fluorocarbon oil FC-40, take 15 μ L, and the second oily liquid (light oil) is a mixed solution of benzylpolysiloxane PD5 and polysorbate additive (Span 80), wherein Span The volume of 80 is 5% of the volume of ...

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Abstract

The invention provides a biochemical micro-reaction system which comprises a first oily liquid, a second oily liquid, a water-based gene sample and a reaction reagent, wherein the density of the first oily liquid is greater than that of the second oily liquid; the water-based gene sample and the reaction reagent are embedded between the first oily liquid and the second oily liquid to form a sandwich structure; the water-based gene sample and the reaction reagent are immiscible and do not react with the first oily liquid and the second oily liquid; and the first oily liquid and the second oily liquid are immiscible. In the biochemical micro-reaction system, each independent sample in a biochemical reaction process can be completely controlled and separated to avoid cross contamination, and a large quantity of consumables can be saved. The invention also provides a high-throughput sequencing database builder based on the biochemical micro-reaction system and an application.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a biochemical micro-reaction system, a high-throughput sequencing library building instrument and its application. Background technique [0002] At present, the mainstream of the DNA sequencer market is the second-generation high-throughput sequencing platform, mainly including Illumina2000 / 2500 / 4000, Roche's 454 sequencer, and ABI's SOLID sequencer. , all require the preparation of sequencing libraries. The process of library construction (library preparation) involves the addition of gene samples, the addition of buffer and enzyme reagent mixture, PCR (polymerase chain reaction), magnetic bead purification, etc. The process is complex and the liquid system used is huge, resulting in mixed samples. and high risk of sample contamination. [0003] In order to solve the above difficulties, there are two existing automatic library preparation solutions as follows: one is based on ...

Claims

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Application Information

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IPC IPC(8): C40B50/06C12M1/38C12M1/36C12M1/34
CPCC12Q1/6869C12Q2563/159C12Q2535/122C12Q2563/143
Inventor 李星军
Owner 李星军
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