Kit for testing schizophrenia related gene and preparation method thereof
A technology for schizophrenia and kits, which is applied in the field of kits for detecting schizophrenia-associated genes and its preparation, and can solve the problems of high cost, high price, and long cycle
Inactive Publication Date: 2011-03-16
SHANGHAI JIAO TONG UNIV
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Problems solved by technology
TaqMan probes need to be customized from AB company, the cycle is longer and the price is more expensive
Although the results of sequencing technology are more reliable, but for the SNP typing of specific loci, this method is relatively expensive and takes a long time
Method used
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Embodiment 1
[0026] 1. Blood Sample Collection and Total DNA Extraction
[0027] The peripheral blood of volunteers was collected to prepare total DNA. DNA was extracted directly using Qiagen's DNA extraction kit, and the extraction method was carried out according to the instructions of the kit. After extraction, use NanoDrop to detect the concentration of each DNA sample, then adjust each sample to 10ng / ul, and store it at -20 degrees for later use.
[0028] 2. PCR reaction
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Abstract
The invention relates to a kit for testing a schizophrenia related gene and a preparation method thereof in the technical field of molecular biology. The kit comprises a PCR reagent group and a LDR reagent group, wherein the PCR reagent group comprises buffer solution, dNTP mixed solution, Taq DNA polymerase, pure water, and four pairs of amplification primers special for PCR, and the LDR reagent group comprises buffer solution, dNTP mixed solution, Taq DNA ligase, pure water, and four pairs of probes special for LDR. The method comprises the steps of: designing and synthesizing PCR primers by using the primer design software PRIMER3, choosing a TM value thereof about 60 degrees, designing and synthesizing the LDR probe which comprises two 5'end probes and a 3'end probe on each locus, and using TAKARA products or New England Biolabs products as the Taq DNA polymerase and the Taq DNA ligase. The invention can realize rapid, efficient and accurate parting for the specific SNP locus in the schizophrenia related gene.
Description
technical field [0001] The invention relates to a kit in the field of molecular biology technology and a preparation method thereof, in particular to a kit for detecting schizophrenia-associated genes and a preparation method thereof. Background technique [0002] Ligase can connect two oligonucleotide fragments that are completely complementary to the target sequence to form a longer oligonucleotide probe. If they are not completely complementary, then the ligation reaction will not take place and no longer oligonucleotide probes will be produced. According to this principle, the genotype of the SNP site can be detected by designing oligonucleotide probes of different lengths according to the genotype of the SNP site. According to the length classification of the final product, the genotype of the SNP site can be determined, which is the ligase detection reaction (Ligase Detection Reaction). [0003] The human genome is the ultimate carrier of genetic information, and the...
Claims
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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 师咏勇李涛李志强冯国鄞贺林
Owner SHANGHAI JIAO TONG UNIV
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