Primer and kit for detecting septin9 gene methylation
A methylation and kit technology, applied in the field of gene septin9 methylation detection, can solve the problems of low detection sensitivity and specificity, and achieve the effects of increased sensitivity and specificity, high sensitivity and easy interpretation
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0035] Embodiment 1 kit preparation
[0036] Primer and probe sequences contained in the kit:
[0037] Septin9 forward primer F: 5'-CCGAAATAATCCCATCCAAC-3',
[0038] Septin9 reverse primer R: 5'-GCGATTCGTTGTTTATTAGTTATTAT-3';
[0039] Internal reference gene forward primer F: 5'-GTGATGGAGGAGGTTTAG-3',
[0040] Internal reference gene reverse primer R: 5'-AAATTACAAAAACCACAA-3';
[0041] septin9 gene probe: FAM-TTATGTCGGATTTCGCGGTTAA-MGB-BHQ1;
[0042] Internal reference gene probe: VIC-CACCACCCAAACACACAATAACAAAC-BHQ1;
[0043] Blocker1 primer: 5'-ACCCGCTGCCCACCAGCCATCATG-CY3-3';
[0044] Blocker2 primer: 5'-TATGTTGGATTTTGTGGTTAATGTGTA-CY3-3'.
[0045] (1) Premixed primer-probe mixture for detecting septin9 gene methylation:
[0046]
[0047] (2) DNA PCR amplification MIX:
[0048]
Embodiment 2
[0049] Embodiment 2 kit detects septin9 gene methylation
[0050] 1) 10 cases of plasma samples were taken for free DNA extraction, and the free DNA extraction kit was from Tiangen Biochemical Technology Co., Ltd.;
[0051] 2) The extraction of free DNA is completed, and the extracted DNA is converted to bisulfite to obtain purified DNA. The conversion kit is from Tiangen Biochemical Technology Co., Ltd.;
[0052] 3) Prepare PCR amplification reaction system:
[0053]
[0054]
[0055] 4) The PCR amplification procedure is as follows:
[0056] The first amplification stage: UDG enzyme reaction at 37°C for 2 minutes;
[0057] The second amplification stage: pre-denaturation at 95°C for 3 minutes;
[0058] The third amplification stage: denaturation at 94°C for 15s, annealing and extension at 60°C for 35s, 45 cycles;
[0059] Signal collection, in the third stage, FAM and VIC signals were collected at 60°C.
[0060] 5) Analysis of test results
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