Novel closed type nucleic acid visual detecting method for coupling nucleic acid amplification reaction, nucleic acid intrusive reaction and nano-particle chromogenic reaction
A nucleic acid invasion reaction and nucleic acid amplification reaction technology, applied in the field of molecular biology, can solve the problems of cross-contamination of amplification products, low sensitivity and low discrimination, and achieve the effects of low cost, high sensitivity and high specificity
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0062] Example 1 A single-tube closed-tube visual nucleic acid detection method using nucleic acid amplification reaction combined with cascade nucleic acid invasion reaction and nano-gold chromogenic detection to detect ultraviolet quantitative genomic DNA.
[0063] The whole genome DNA is used as a template for detection after gradient dilution to examine the sensitivity of the method of the present invention.
[0064] Reaction conditions:
[0065] The reaction system composition of this step is 10mM Tris-HCl (pH7.5), 0.05%Tween-20, 0.05%Nonidet P40, 4mM MgCl 2 , 30mM NaCl, 0.5μM upstream primer (sequence: 5'-CCG GGA GTT GGG CGA GTA C-3', Tm value is 70.8°C, SEQ ID NO.1), 0.5μM downstream primer (sequence: 5'- GCC CCC AGC AGG TCC C-3', Tm value is 71.8°C, SEQ ID NO.2), 0.2 μM upstream probe (sequence: 5'-CTC TGC CAC CAG CTC CCA-3', Tm value is 68.5°C , SEQ ID NO.3), 0.2 μM downstream probe (the wild-type probe sequence is: 5'-CGC GCC GAG GCG AAC TTG GGC GAG-C 3 -3', Tm va...
Embodiment 2
[0067] Example 2 A single-tube closed-tube visual nucleic acid detection method using nucleic acid amplification reaction combined with cascade nucleic acid invasion reaction and nanoprobe chromogenic detection to detect SNP site typing.
[0068]Using the present invention to detect SNP site typing needs to design corresponding downstream probes for different types of detection sites, and divide them into two tubes for detection. When the reaction system contains a template that matches the downstream probe, hybridize with the nanoprobe When the reaction system does not contain a template that matches the downstream probe, it will aggregate after hybridizing with the nanoprobe, thereby realizing the typing of the SNP site.
[0069] Reaction conditions:
[0070] The reaction system composition of this step is 10mM Tris-HCl (pH7.5), 0.05%Tween-20, 0.05%Nonidet P40, 4mM MgCl 2 , 30mM NaCl, 0.5μM upstream primer (sequence: 5'-CCG GGA GTT GGG CGA GTA C-3', Tm value is 70.8°C, SEQ ...
Embodiment 3
[0072] Example 3 Nucleic acid amplification reaction combined with cascaded nucleic acid invasion reaction and nano-gold chromogenic detection single-tube closed-tube visualized effect of different storage time on gold nano-precipitation after nucleic acid detection.
[0073] The whole genome DNA is used as a template for detection, and after the detection reaction is completed, it is placed at room temperature for different times to investigate the precipitation of the gold nanometer aggregation reaction in the method of the present invention.
[0074] Reaction conditions:
[0075] The reaction system composition of this step is 10mM Tris-HCl (pH7.5), 0.05%Tween-20, 0.05%Nonidet P40, 4mM MgCl 2 , 30mM NaCl, 0.5μM upstream primer (sequence: 5'-CCG GGA GTT GGG CGA GTA C-3', Tm value is 70.8°C, SEQ ID NO.1), 0.5μM downstream primer (sequence: 5'- GCC CCC AGC AGG TCC C-3', Tm value is 71.8°C, SEQ ID NO.2), 0.2 μM upstream probe (sequence: 5'-CTC TGC CAC CAG CTC CCA-3', Tm value ...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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