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Polyacrylamide derivative modified filter paper and application thereof in nucleic acid separation and enrichment

A technology of polyacrylamide and derivatives, applied in filtration separation, filter paper, separation methods, etc., can solve the problems of high-efficiency enrichment and low enrichment rate of difficult nucleic acids, achieve high-efficiency separation and enrichment, improve sensitivity, and simple operation Effect

Active Publication Date: 2021-06-18
HANGZHOU ZIJING BIOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods still have the disadvantages of introducing chloroform and ethanol and other interfering substances that can inhibit subsequent amplification during the extraction process, the enrichment rate is low, and it is difficult to adapt to the high-efficiency enrichment of nucleic acids in large-volume samples.

Method used

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  • Polyacrylamide derivative modified filter paper and application thereof in nucleic acid separation and enrichment
  • Polyacrylamide derivative modified filter paper and application thereof in nucleic acid separation and enrichment
  • Polyacrylamide derivative modified filter paper and application thereof in nucleic acid separation and enrichment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1, Extraction of New Crown Virus RNA Joint QPCR Detection

[0061] First, the preparation of acrylamide derivatives modified filter paper

[0062] (1) Cleaning and activation of filter paper surface

[0063] The quartz matrix fiber filter was placed in a 1 M hydrochloric acid solution to remove impurities of the surface of the filter paper fiber. After cleaning and activation, use a large amount of deionized water to clean, so that the pH recovery of the filter paper is unknown.

[0064] (2) Siliconlated modification of filter paper

[0065] The activated filter paper is placed in a 1: 1 mixture of ethanol containing 0.1% by weight of 3- (methacryloxyoxylated) propyltrimethoxysilane and water, and the reaction is used after 1 hour under room temperature. Clean three times, after cleaning, then dried on the hot plate of 80 ° C.

[0066] (3) polymerization reaction of acrylamide derivative monomer

[0067] The reaction premix is ​​configured before the polymerization ...

Embodiment 2

[0093] Example 2, Extraction of the Chlamydia Terminal DNA and LAMP Amplifies.

[0094] First, the preparation of acrylamide derivatives modified filter paper

[0095] (1) Cleaning and activation of filter paper surface

[0096] The glass matrix fiber filter was placed in a 1 M hydrochloric acid solution to remove impurities of the surface of the filter paper fiber. After cleaning and activation, use a large amount of deionized water to clean, so that the pH recovery of the filter paper is unknown.

[0097] (2) Siliconlated modification of filter paper

[0098] The activated filter paper was placed in an ethanol containing 0.1% by weight of 3- (methylpropyleneoxyoxylated) propyltrimethoxysilane with water 1: 1 common mixture, after 1 h under room temperature, use deionized water cleaning. Three times, after cleaning, place it on the hot plate of 80 ° C.

[0099] (3) polymerization reaction of acrylamide derivative monomer

[0100] The reaction premix was configured before the poly...

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Abstract

The invention discloses polyacrylamide derivative modified filter paper and application thereof in nucleic acid separation and enrichment. The polyacrylamide derivative modified filter paper is fiber filter paper modified with a modifier, and the modifier is a polymer of one or more acrylic acid derivative monomers. The invention provides a nucleic acid extraction method based on the polyacrylamide derivative modified filter paper, the method can be used for DNA separation and enrichment and also can be used for RNA separation and enrichment, the polyacrylamide derivative modified filter paper serves as a core, the method comprisesa DNA extraction reagent, an RNA extraction reagent and a device, and the reagents mainly comprise a lysis solution, a cleaning solution and a nucleic acid releasing agent. The device consists of a nucleic acid extraction column, a filter paper fixing bottom support, a centrifugal tube and a PCR tube, comprises four basic steps of cracking, nucleic acid capturing, cleaning and filter paper transferring, and has the advantages of simplicity in operation, capability of being used for separating nucleic acid in a large-volume biological sample, no organic solvent such as ethanol, easiness in integration on a micro-fluidic chip and high enrichment rate.

Description

Technical field [0001] The present invention relates to a polyacrylamide derivative modification filter paper and its application in nucleic acid separation, belonging to the field of nucleic acid extraction. Background technique [0002] The extraction of nucleic acids is widely used in the detection of infectious diseases, diagnosis of tumors, prenatal screening, forensic identification, food safety testing, molecular biology research. The new coronavirus epidemic in the end of 2019 fully showed the huge role of nucleic acid detection in the prevention and control of epidemic, and also exposed many problems in nucleic acid detection. For example, it is limited by nucleic acid detection capabilities, and the screening of low-risk populations is difficult to launch a large area. The detection of multiple samples can be greatly improved, and it can also reduce screening costs, which is a program that expands the nucleic acid screening capacity of infectious diseases. However, afte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): D21H13/40D21H17/55D21H21/14D21H27/08
CPCD21H27/08D21H13/40D21H17/55D21H21/14Y02A50/30
Inventor 刘鹏李尚霖李保韩冰倩
Owner HANGZHOU ZIJING BIOLOGY CO LTD
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