Specific detection primers and detection liquid phase chip for FYCO1 gene mutation
A technology of specificity and detection solution, which is applied in the field of molecular biology, can solve the problems of time-consuming and labor-intensive sequencing reactions, high false positive rate, and insufficient flexibility, so as to avoid uncertain factors, good detection specificity, and avoid cross-reactions Effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0026] Embodiment 1 FYCO1 gene mutation detection liquid chip mainly includes:
[0027] 1. ASPE Primers
[0028] Specific primer sequences were designed for the wild-type and mutant types of two common genotypes A92G and C57T of the FYCO1 gene. ASPE primers consist of "tag sequence + specific primer sequence". ASPE primer sequences are shown in the table below:
[0029] Table 1 ASPE primer sequence (specific primer sequence) of FYCO1 gene
[0030]
[0031]
[0032] Table 2 ASPE primer sequence (tag sequence) of FYCO1 gene
[0033]
[0034] Each ASPE primer includes two parts, the 5' end is the specific tag sequence for the anti-tag sequence on the corresponding microsphere (as shown in the above table 2), and the 3' end is the mutant or wild type specific primer fragment ( as shown in Table 1 above). All ASPE primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. Each synthesized primer was prepared into a 100 pmol / mL stock solut...
Embodiment 2
[0048] Example 2 Detection of samples using the FYCO1 gene mutation detection liquid chip described in Example 1
[0049] The formula of described various solutions is as follows:
[0050] 50mM MES buffer (pH5.0) formula (250ml):
[0051]
[0052] 2×Tm hybridization buffer
[0053] Reagent
source
Final concentration
Dosage per 250ml
1M Tris-HCl, pH8.0
SigmaT3038
0.2M
50ml
5M NaCl
Sigma S5150
0.4M
20ml
Triton X-100
Sigma T8787
0.16%
0.4ml
[0054] Store at 4°C after filtration.
[0055] ExoSAP-IT kit was purchased from US USB Company.
[0056] Biotin-labeled dCTP was purchased from Shanghai Sangon Bioengineering Technology Service Co., Ltd.
[0057] 1. Sample DNA extraction:
[0058] Refer to the relevant methods of DNA extraction in "Molecular Cloning" to obtain the DNA to be detected.
[0059] 2. PCR amplification of samples to be tested
[0060] Design 2 pair...
Embodiment 3
[0104] The liquid phase chip of embodiment 3 different ASPE primers detects the SNP site of FYCO1 gene
[0105] 1. Design of liquid phase chip preparation (selection of Tag sequence and Anti-Tag sequence)
[0106] Taking the FYCO1 gene A92G and C57T site mutation detection liquid chip as an example, the specific primer sequence at the 3' end of the ASPE primer was designed for the wild type and mutant type of A92G and C57T, and the Tag sequence at the 5' end of the ASPE primer was selected from SEQ ID NO.5-SEQ ID NO.10, correspondingly, the anti-tag sequence coated on the microsphere and complementary to the corresponding tag sequence is selected from SEQ ID NO.11-SEQ ID NO.16. The specific design is shown in the following table (Table 9). The synthesis of ASPE primers, microspheres coated with anti-tag sequences, amplification primers, detection methods, etc. are as described in Example 1 and Example 2.
[0107] Table 9 Design of liquid phase chip preparation
[0108]
...
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