Method and kit for detecting multiple cancer risk susceptibility
A technology for detection kits and reagents, applied in the field of tumor molecular genetics, can solve the problems of high cost and inappropriateness, and achieve the effects of good sensitivity and improved discrimination efficiency.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0029] Example 1: Preliminary studies have found that 3q24-26 is closely linked to multi-cancer families
[0030] In the previous research, the inventor discovered a rare multi-cancer family in Hunan, with more than 103 people in six generations. Among them, there were 15 patients, and 10 survived, involving various benign and malignant tumors. For the family map of multiple cancers, please refer to the attached figure 2. In order to locate the chromosome segment linked to the disease, we first used a high-throughput SNP chip (Human Mapping 500K SNP Array of Affymetrix Company) to scan and type the whole genome, and performed single-point parameter linkage analysis through Merlin software. The area with LOD value greater than 2 is obtained. Later, microsatellites were used for fine positioning in this area. When multi-point non-parametric linkage analysis was performed by Genehunter software, the maximum LOD value reached 10.674 (p=0.00195), which was located between D3S158...
Embodiment 2
[0037] Example 2: Preparation of tumor risk susceptibility SNP diagnostic kit
[0038] (1) Primer design for SNP loci
[0039] 1. Selection of template sequence
[0040] The template sequence was from the UCSC database ( http: / / genome.ucsc.edu / ), select 300 bp upstream and downstream of the SNP site, to ensure that the PCR product is in a suitable fragment range for easy detection.
[0041] 2. Primer Design
[0042] There are 3 basic principles for PCR primer design: firstly, primers should be closely combined with the template, secondly, there should be no stable dimer or hairpin structure between primers, and thirdly, primers should not cause DNA polymerization at other non-target sites (i.e. mismatch). Specific considerations include: primer length, product length, sequence Tm value (melting temperature), ΔG value (internal stability), primer dimer and hairpin structure (duplex formation and hairpin), error triggering Site (false priming site), primer and product GC ...
Embodiment 3
[0074] Embodiment 3: the use steps of SNP diagnostic kit
[0075] Please refer to the attached figure 1 .
[0076] Step 1 Blood sample collection and gDNA extraction
[0077] Collection and processing of blood samples: using the German Greiner company (Non-replaceable) 5ml EDTA anticoagulant tube, collect 5ml of peripheral blood sample from the individual to be tested, store at 4°C, and extract gDNA within two weeks. If the gDNA is not extracted immediately, the anticoagulant tube can be stored in the refrigerator at -20°C, and extracted together after collecting multiple samples.
[0078] gDNA extraction: Blood gDNA was extracted using TaKaRa Company Whole Blood Genome Extraction Kit (Universal GenomicDNA Extraction Kit Ver.3.0) (see the product manual for details). Take 2 μl of DNA sample to run on the gel, and take 2 μl of gDNA sample to dilute to 100 μl at the same time, measure the absorbance value, and calculate the extracted DNA concentration according to the formu...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More