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Single cell transcriptome sequencing library construction platform and use method thereof

A transcriptome sequencing and library construction technology, applied in the field of single-cell transcriptome sequencing library construction platform, can solve the problems of incapability of cell capture and detection, purification and library integration, difficulty in providing a fully automatic integrated experimental platform, etc., to achieve Complicated operation, overcoming pump valve operation and flow rate control, high damage effect

Pending Publication Date: 2022-08-05
厦门德运芯准科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing DMF chip still cannot integrate cell capture with detection, purification and library construction, and it is difficult to provide a fully automatic integrated experimental platform

Method used

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  • Single cell transcriptome sequencing library construction platform and use method thereof
  • Single cell transcriptome sequencing library construction platform and use method thereof
  • Single cell transcriptome sequencing library construction platform and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Reference below Figure 1 to Figure 5 Describe in detail the single-cell transcriptome sequencing library construction platform according to the present invention, which includes a digital microfluidic chip 100 , a circuit control module 200 , a controller 300 and a magnetic separation control module 400 .

[0077] The magnetic separation control module 400 is located at the lower part of the digital microfluidic chip 100 for generating a magnetic field, so as to facilitate DNA purification using magnetic beads on the platform. The magnetic separation control module 400 includes a magnet 410 and a cam 411 for controlling the up and down movement of the magnet.

[0078] The digital microfluidic chip 100 also includes a lower electrode plate 120 and an upper electrode plate 110 which are arranged in parallel and opposite to each other, and are separated by space for liquid movement; the lower electrode plate 120 sequentially includes a bottom plate 121, an electrode layer...

Embodiment 2

[0116] 1. Add the cell suspension, lysate and reverse transcription reagents to the storage area 2 first, and pre-energize in the corresponding storage area.

[0117] Lysate Reaction Solution Recipe

[0118]

[0119] Reverse Transcription Reaction Solution Recipe

[0120]

[0121]

[0122] 2. The electrode drive circuit is controlled by the controller 300 and the circuit control module 200, and the on-off control of the electrodes is carried out in a preset order, so that a droplet of cell suspension is generated from the electrode unit of the liquid storage area 2, and the The drive electrode array moves this droplet to the area of ​​hydrophilic sites 124A. This process is repeated so that each hydrophilic site area is loaded with droplets of cell suspension.

[0123] 3. Adjust the position of the cells in the droplet by applying a voltage to the adjacent electrodes so that a single cell is located above the hydrophilic site. Power off the drive electrodes at the ...

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Abstract

The invention relates to a single-cell transcriptome sequencing library construction platform which comprises a digital micro-fluidic chip, a circuit control module, a controller, a magnetic separation control module of a magnet and a cam located at the bottom of the magnet and used for adjusting the magnet to ascend and descend. The chip comprises an upper pole plate and a lower pole plate, the lower pole plate comprises a substrate, an electrode layer, a dielectric layer and a hydrophobic layer, and the upper pole plate comprises a hydrophobic layer, a grounding conductive layer and a top plate; the lower stage plate is divided into a liquid drop driving area and a plurality of liquid storage areas; the hydrophobic layer comprises hydrophilic points; the electrode layer comprises a driving electrode array and a plurality of liquid storage electrode arrays. The invention also relates to a use method of the library construction platform. According to the construction platform and the use method, full-automatic reagent control and addition, flexible and rapid capture of single cells and sensitive, accurate and stable gene detection are realized, and the cost is low. According to the integrated preparation platform, the experimental mode and steps are greatly simplified, and high-throughput cell genetic information detection and analysis become possible.

Description

technical field [0001] The invention relates to the field of digital microfluidic chips, in particular to a single-cell transcriptome sequencing library construction platform and a method for using the same. Background technique [0002] Cells are the basic units of living organisms. From the point of view of a single organism, the genes in each cell are basically the same, but their gene expression shows great heterogeneity, which constitutes the diversity of tissues, organs and systems. This cell-to-cell heterogeneity occurs even within cells of the same type. Differences in gene expression between cells have crucial implications for biological processes such as embryonic development, cancer growth, immune responses, and more. Population-level transcriptome analysis techniques cannot reveal information about cell-to-cell heterogeneity, so it is necessary to develop single-cell transcriptome analysis techniques. [0003] Based on the second-generation sequencing technolo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/42C12M1/36C12M1/38C12M1/34C12M1/33C12M1/02C12M1/00C12Q1/6806C40B60/14C40B50/06B01L3/00
CPCC12Q1/6806C40B60/14C40B50/06B01L3/5027B01L3/50273B01L2200/10B01L2300/0861B01L2300/0887B01L2300/165B01L2400/0415C12Q2525/191C12Q2531/113C12Q2563/159C12Q2565/629C12Q2523/308
Inventor 杨朝勇许醒张倩倩杨健蔡林峰
Owner 厦门德运芯准科技有限公司
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