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Method for carrying out a biochemical protocol in continuous flow in a microreactor

a microreactor and biochemical technology, applied in the direction of fluid speed measurement, optical light guide, water/sand/air bath, etc., can solve the problems of not allowing large-scale protocols to be performed, affecting the flow rate of the microreactor, so as to reduce the number of distributions, improve the throughput, and reduce the cost

Inactive Publication Date: 2005-12-22
SERONO GENETICS INST SA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a device and method for conducting biochemical or chemical processes on samples using a microfluidic substrate with at least one microchannel for carrying out reactions or sequences of reactions. The device includes a microfluidic substrate with at least one microchannel and a thermal support for controlling the temperature in different zones of the channel. The device can be used with a continuous flow of samples and reagents, allowing for a high throughput and reducing the number of distributions to be carried out. The device is semi-disposable and can be easily added and removed from the thermal support. The method involves feeding a continuous flow of solution containing the sample and reagents, injecting the reagents, and transferring heat between the thermal support and the temperature-regulated portion of the channel. The device and method provide a cost-effective and efficient way to conduct biochemical or chemical processes.

Problems solved by technology

This device does not, however, integrate the mixing of reagents, and it does not allow large scale protocols to be performed.
One of the difficulties in implementing these devices resides in the movement of the fluids.
These systems which integrate electrodes, microvalves or micropumps are very costly and their complexity does not allow large scale applications for simultaneously treating a very large number of samples.
One of the major difficulties is the distribution, mixing and transport of a very large number of products in parallel or in series.

Method used

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  • Method for carrying out a biochemical protocol in continuous flow in a microreactor
  • Method for carrying out a biochemical protocol in continuous flow in a microreactor
  • Method for carrying out a biochemical protocol in continuous flow in a microreactor

Examples

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example 1

PCR Reaction in a Continuous Flow in a Microfluidic Device

[0370] A PCR reaction mixture was run through a channel in which a PCR was performed.

[0371] The microfluidic substrate comprised 10 channels in parallel chemically etched in silicone. The channels were rectilinear with a diameter of the order of about 600 μm in the reaction zones. The surface area of the section of a channel was of the order of about 0.25 mm2.

[0372] The PCR was carried out in parallel in 3 channels. The volume of one PCR reaction was 1.2 μl, but ten identical PCR reactions (12 μl (10×1.2 μl)) were performed for out of the device post PCR sample analysis (quantification and size analysis).

[0373] The microfluidic substrate was siliconized just before the substrate was used. Before the PCR reaction, all the channels were filled with previously degassed and filtered water. A high flow rate of the order of 25 μl / min was applied for 15 minutes to remove all the air bubbles present in the circuits. A previously ...

example 2

Integration of a Genotyping Protocol in a Continuous Flow Microfluidic Device

[0379] In one embodiment, the reaction mixture runs through a channel in which all the steps required for a genotyping protocol are performed: PCR, purification, microsequencing reaction and detection.

[0380] The microfluidic substrate comprises 100 channels in parallel. These channels are rectilinear and have a diameter of the order of 600 μm in the reaction zones. The surface area of the section of a channel is of the order of 0.25 mm2.

[0381] The genotyping protocol is carried out in parallel in the 100 channels. 100 samples are injected in parallel into the channels, and 100 such injections of sample are carried out sequentially, so that each channel contains 100 injections of the same sample. The microfluidic substrate thus makes it possible, by means of a cross-distribution of 100 samples for 100 polymorphisms, to carry out 10,000 genotyping reactions on a microfluidic substrate.

[0382] The injection...

example 3

Genotyping by the Method of Allele-Specific Ligase Chain Reaction (LCR)

[0399] Allele-specific LCR, as disclosed in Barany et al. (PCR Meth. Appl. 1: 5-16 (1991), the contents of which are incorporated herein by reference in its entirety), employs four oligonucleotides two of which hybridize to one strand of target DNA and a complementary set of adjacent oligonucleotides, which hybridize to the opposite strand. Thermostable DNA ligase will covalently link each set, provided there is complete complementarity at the junction. A single-base mismatch at the oligonucleotide junction will not be amplified and is therefore distinguished; a second set of mutant-specific oligonucleotides is used in a separate reaction to detect or confirm the mutant allele(s).

[0400] A homogeneous phase protocol for allele-specific LCR can be carried out in accordance with the present invention by introducing into each channel 10 μl of starting mixture comprising: the DNA with the target sequence to be analy...

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Abstract

Devices and methods for carrying out a chemical or biochemical protocol are disclosed. In one embodiment, the chemical or biochemical protocol is performed by cycling at least one thermal transfer member between at least two temperatures while liquid samples on which the chemical or biochemical protocol is to be performed are continuously moving through at least one temperature regulated zone upon which the at least one thermal transfer member acts. In some embodiments, the device comprises a sample transport member that comprises liquid samples in sample receiving regions. The sample transport member moves the samples continuously through a temperature regulated zone which cycles between at least two temperatures while the liquid samples are moving through a temperature regulated zone on which at least one thermal transfer member acts. In some embodiments, the sample receiving regions comprise wells, hydrophillic films or hydrophillic filaments.

Description

RELATED APPLICATIONS [0001] The present application is a continuation of U.S. patent application Ser. No. 09 / 772,280, filed Jan. 29, 2001, which is a Continuation-In-Part application of U.S. patent application Ser. No. 09 / 627,647, filed Jul. 28, 2000, which claims priority to French patent application Serial No. 99 / 09806, filed Jul. 28, 1999; French patent application Serial No. 99 / 11652, filed Sep. 17, 1999; and French patent application Serial No. 99 / 12317, filed Oct. 1, 1999, the disclosures of all of which are incorporated herein by reference in their entireties. BACKGROUND [0002] Microfluidics consist of using microchannels instead of test tubes or microplates to carry out analyses and reactions. These microchannels or microcircuits are etched into silicon, quartz, glass, ceramics or plastic. The size of these channels is on the order of micrometers, while the reaction volumes are on the order of nanoliters or microliters. The principle of a microfluidic device is to guide reac...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): B01J19/00B01L3/00G01N33/50B01L7/00B01L7/02B81B1/00B81C1/00C12M1/00C12M1/22C12M1/34C12M1/38C12N15/09C12P19/34C12Q1/68G01N1/28G01N35/02G01N35/08G01N37/00
CPCB01L3/5025G01N35/08B01L3/502707B01L3/502746B01L3/502784B01L7/02B01L7/52B01L7/525B01L7/5255B01L2200/0673B01L2200/10B01L2300/0816B01L2300/0845B01L2300/0867B01L2300/087B01L2300/1822B01L2300/1827B01L2300/1838B01L2300/1883B01L2400/0415B01L2400/0487F28F2260/02B01L3/5027
Inventor FOUILLET, YVESVAUCHIER, CLAUDECLERC, JEAN-FREDERICPEPONNET, CHRISTINECLAUSTRE, PATRICIACHARLES, RAYMONDSARRUT, NICOLAS
Owner SERONO GENETICS INST SA
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