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Spotting solution for fixing nucleic acid probe, preparation method of spotting solution, and method for fixing nucleic acid probe

A nucleic acid probe and spotting technology, applied in the field of nucleic acid chips, can solve the problems of high material cost, long time consumption, inconvenient operation, etc., and achieve the effect of meeting the requirements of large-scale production, fixing time-saving and low-cost.

Active Publication Date: 2017-05-24
GENETEL PHARMA SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of this process method are: cumbersome steps, long time-consuming, high material cost, inconvenient operation, difficult to meet the requirements of mass production, etc.

Method used

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  • Spotting solution for fixing nucleic acid probe, preparation method of spotting solution, and method for fixing nucleic acid probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] 1. Weigh 0.2g of EDC and add it to 5ml of sunset yellow pigment solution containing 0.1%;

[0100] 2. Mix 80 μL of this solution with probe working solution F1 in equal volume, and centrifuge at 10,000 rpm for 5 min;

[0101] 3. Add the mixture to the sample plate;

[0102] 4. Spread the untreated nylon membrane on the spotter stage;

[0103] 5. Edit the spotting program and start spotting;

[0104] 6. Use the reverse complementary sequence of the F1 probe to hybridize with the chip to record the INS value and negative and positive.

Embodiment 2

[0106] 1. Weigh 0.2g of EDC and add it to 5ml of sunset yellow pigment solution containing 0.5%;

[0107] 2. Mix 80 μL of this solution with probe working solution F1 in equal volume, and centrifuge at 10,000 rpm for 5 min;

[0108] 3. Add the mixture to the sample plate;

[0109] 4. Spread the untreated nylon membrane on the spotter stage;

[0110] 5. Edit the spotting program and start spotting;

[0111] 6. Use the reverse complementary sequence of the F1 probe to hybridize with the chip to record the INS value and negative and positive.

Embodiment 3

[0113] 1. Weigh 0.2g of EDC and add it to 5ml of sunset yellow pigment solution containing 1%;

[0114] 2. Mix 80 μL of this solution with probe working solution F1 in equal volume, and centrifuge at 10,000 rpm for 5 min;

[0115] 3. Add the mixture to the sample plate;

[0116] 4. Spread the untreated nylon membrane on the spotter stage;

[0117] 5. Edit the spotting program and start spotting;

[0118] 6. Use the reverse complementary sequence of the F1 probe to hybridize with the chip to record the INS value and negative and positive.

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Abstract

The invention discloses a spotting solution for fixing a nucleic acid probe, a preparation method of the spotting solution, and a method for fixing the nucleic acid probe. The spotting solution contains 1-(3-dimethylamino propyl)-3-ethyl carbodiimide (EDC) and sunset yellow pigment. During use of the spotting solution provided by the invention, a nylon membrane does not need to be treated by hydrochloric acid and EDC in advance, the spotting solution and a nucleic acid solution are mixed to be directly dripped to the nylon membrane which is not treated to finish fixation of the nucleic acid probe. Therefore, the spotting solution provided by the invention enables fixation of the nucleic acid probe to be time-saving, low in cost and easy to operate, and can meet the large-scale production requirement.

Description

technical field [0001] The present invention relates to the technical field of nucleic acid chips, in particular to a spotting solution for immobilizing nucleic acid probes, a preparation method thereof, and a nucleic acid probe immobilization method. Background technique [0002] In the process of making a nucleic acid chip based on a nylon membrane solid phase carrier, a key step is to immobilize the nucleic acid probe on the nylon membrane. The traditional process method currently adopted is to first use hydrochloric acid, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) to activate nylon membrane (Zhang Yong, Coyne Mazie Y, WillStephen G, et al (1991), Single-base mutational analysis of cancer and genetic diseases using membrane bound modified oligonucleotides [J]. Nucl Acids Res, 19(14): 3929-3933.; Chehab FF and Wall J (1992), Detection of multiple cysticfibrosis mutations by reverse dot blot hybridization:a technology for carrierscreening[J].Human Genetics, 89(2):...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C40B50/18C12Q1/68C40B40/06
Inventor 赵雷李勐黄明辉
Owner GENETEL PHARMA SHENZHEN