Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Aptamer screening method of no-fixed point target substance

A screening method and unfixed technology, applied in the direction of biochemical equipment and methods, microbe determination/inspection, etc., to achieve the effect of strong applicability, convenient operation and low cost

Active Publication Date: 2013-05-22
SHANGHAI JIAO TONG UNIV
View PDF2 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the premise of using this method is that the target substance has a fixed point, so there are few reports on the screening of aptamers without a fixed point.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Aptamer screening method of no-fixed point target substance
  • Aptamer screening method of no-fixed point target substance
  • Aptamer screening method of no-fixed point target substance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Such as figure 1 As shown, this embodiment includes the following steps:

[0034] Step 1) Construct a random oligonucleotide library with a total length of 68bp. The sequence of the oligonucleotide library is: 5'-ACCGACCGTGCTGGACTCT-N30-AGTATGAGCGAGCGTTGCG-3', wherein both ends are fixed sequences of 19bp, and the middle 30 bases The basis is a random sequence.

[0035] Step 2) design three primers, wherein P1 (5'-ACCGACCGTGCTGGACTCT-3') and P2 (5'-CGCAACGCTCGCTCATACT-3') are used as the upstream primer and downstream primer of PCR amplification oligonucleotide respectively, and P3 (5' -Biotin-CGCAACGCTCGCT CATACT-3') was used to immobilize the oligonucleotide library on streptavidin-labeled agarose particles.

[0036] Step 3) Take a random oligonucleotide library, mix it with the P3 sequence uniformly at a volume ratio of 1:2-3, and anneal the mixture. The annealing conditions are: denaturation at 94°C for 60 seconds, annealing at 59°C for 60 minutes, and 25°C Exten...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses an aptamer screening method of a no-fixed point target substance in the technical field of nucleic acid aptamer screening in analytical chemistry. The method comprises the following steps of: screening in a systematic evolution of ligands by an exponential enrichment (SELEX) method by building a random oligonucleotides library, and an upstream primer, a downstream primer and a fixed oligonucleotides library primer for polymerase chain reaction (PCR) proliferation and utilizing an agarose particle fixed oligonucleotides library with a streptavidin mark to obtain a PCR product for clone sequencing; and obtaining aptamers bonded with the no-fixed point target substance with the special affinity after the sequencing is finished. The method is applied to screening of heavy metal cadmium ion aptamers, so that 13 aptamers which are bonded with the cadmium ion with the high specificity and high affinity are obtained; and therefore, the aptamer screening method is good in fixing effect, high in adaptability, low in cost and suitable for aptamer screening of the no-fixed point target substance.

Description

technical field [0001] The present invention relates to a method in the technical field of analytical chemistry nucleic acid aptamer screening, in particular to a method for immobilizing an oligonucleotide library using streptavidin-labeled agarose particles, and screening non-fixed-point targets by SELEX technology The method of aptamers of substances. Background technique [0002] SELEX (Systematic Evolution of Ligands by Exponential Enrichment) technology is a new combinatorial chemistry technology developed in the early 1990s. Using this technology, nucleic acid aptamers with specificity and high affinity to target substances can be screened from random single-stranded oligonucleotide libraries. The basic idea is to chemically synthesize a single-stranded oligonucleotide library in vitro, mix it with the target substance, form a target substance-nucleic acid complex, elute unbound nucleic acid, separate the nucleic acid molecule bound to the target substance, and use th...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 周培吴远根詹深山王法泽刘乐邢海波詹学佳
Owner SHANGHAI JIAO TONG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com