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Construction method for high-throughput micro-fluidic chip detecting system

A technology of microfluidic chip and detection system, which is applied in the field of construction of high-throughput microfluidic chip detection system, and can solve the problems that no microfluidic chip has been publicly published.

Inactive Publication Date: 2013-01-30
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] So far, no application of crosslinkers to microfluidic chips has been published

Method used

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  • Construction method for high-throughput micro-fluidic chip detecting system
  • Construction method for high-throughput micro-fluidic chip detecting system
  • Construction method for high-throughput micro-fluidic chip detecting system

Examples

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Embodiment 1

[0094] Example 1, Construction of High-throughput Microfluidic Transgenic Protein Chip

[0095] Step 1: Based on μParaflo TM The microfluidic biochip synthesizes six single-stranded oligonucleotides of different sequences in situ, and the 3' end of the nucleic acid is connected to the bottom of the chip. Number the positions of each probe (i.e. single-stranded oligonucleotide) from A to F, repeat 6 positions for each probe, and number each position, for example, type A probes are counted as A1, A2, ... A5 , A6, other types of probes are similar. Therefore, there are a total of 36 probe sites on the chip. The sequence and number of each probe are as follows in Table 1:

[0096] Numbering Nucleic acid sequence (5'-3')

A 3'CAC ACG CCT TTA CGA AGA CGA T 5'

B 3' CAG GTC AAA AGG GTC CTT AGG GA 5'

C 3'TAG TGT GTT TCC GTT GAA AAC A 5'

D 3'TAG TTG TCT GTA ATT AAC CCG CG 5'

E 3' AAA CCG TTA CCA TCT TGA GTG TGG C 5'

...

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Abstract

The invention discloses a construction method for a high-throughput micro-fluidic chip detecting system. The construction method mainly comprises three processes, namely action of antibody and nucleotide crosslinking and the micro-fluidic chip, action of crosslinked antibody and nucleotide and a micro-fluidic chip, and action of fluorescent second antibodies and the micro-fluidic chip. The antibody and nucleotide crosslinking mainly comprises the following steps of: modifying and desalting antibodies; modifying and desalting nucleotide; and crosslinking and purifying the modified antibodies and the modified nucleotide and the like. The action of the antibody and nucleotide crosslinking and the high-throughput micro-fluidic chip mainly comprises the following steps of: eluting, melting and closing the chip; and reacting and eluting the crosslinking and the chip. The action of the fluorescent second antibodies and the micro-fluidic chip comprises the following steps of: idiosyncratically reacting Cy3 labelled second antibodies and a crosslinking antibody part connected to the chip; and performing subsequent eluting, scanning, melting and the like.

Description

technical field [0001] The invention relates to a construction method of a high-throughput microfluidic chip detection system based on the cross-linking of antibodies and nucleotides. Background technique [0002] Although the study of gene chips has made great progress, there are still reports that the results are not applicable to the expression of real protein traits in the life process. With the introduction of the concept of proteomics, and the fact that protein, as the final form of biological activity, can directly and truly reflect the research results, people need a high-throughput protein analysis technology for in-depth research, and protein chip technology has emerged as the times require. . [0003] As an emerging technology in the field of life sciences, protein chips have been more and more widely used in food hygiene and quarantine, medical diagnosis, and new drug development. In the case of a small amount of biological samples, the protein chip has played ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 郑晓冬殷赟沙莎高晓莲周小川余挺
Owner ZHEJIANG UNIV
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