The use of abcc4 gene polymorphism site rs3742106 and its detection primers and kit
A technology of gene polymorphism and kit, applied in the field of medicine
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Embodiment 1
[0082] A kit and method for predicting the chemotherapy efficacy of colorectal cancer patients based on single base extension-MALDI / TOF / MS method to determine the genotype of ABCC4 gene polymorphism site rs3742106.
[0083] 1. The composition of the kit:
[0084] 1. Primers for detecting the rs3742106 genotype of the ABCC4 gene polymorphism site;
[0085] Primers for amplifying polymorphic loci:
[0086] SEQ ID No. 1: 5'-CAAGATGGTGCAACAACTGG-3'
[0087] SEQ ID No.2: 5'-GTTAGAGCTGGAGATCCTTG-3'
[0088] Primers for single base extension of polymorphic loci:
[0089] SEQ ID No.4: 5'-AGTCCGTTCCGAAGGCATTT-3'
[0090] 2. PCR amplification enzyme and corresponding buffer;
[0091] 3. dNTPs;
[0092] 4. PCR product purification enzyme;
[0093] 5. Single base extension reaction enzyme and corresponding buffer;
[0094] 6. Enzyme purification of single base extension products.
[0095] 2. Genotype detection and curative effect prediction:
[0096] 1. Collection of specimens: ...
Embodiment 2
[0127] A kit and method for determining the genotype of the ABCC4 gene polymorphism site rs3742106 based on DNA sequencing.
[0128] 1. The composition of the kit:
[0129] 1. Primers for amplifying the polymorphic site rs3742106 of the ABCC4 gene:
[0130] SEQ ID No. 1: 5'-CAAGATGGTGCAACAACTGG-3'
[0131] SEQ ID No.2: 5'-GTTAGAGCTGGAGATCCTTG-3'
[0132] 2. PCR amplification enzyme and corresponding buffer;
[0133] 3. dNTPs;
[0134] 4. PCR product purification enzyme.
[0135] 2. Genotype detection
[0136] 1. Collect specimens
[0137] 1ml of whole blood was drawn from the patient's peripheral vein and placed in a 0.5M EDTA anticoagulant tube.
[0138] 2. Genomic DNA extraction
[0139] Use the Genomic DNA Extraction Kit to extract DNA according to the instructions.
[0140] 3. Genotype detection
[0141] (1) Take 1 μl of DNA sample, and use 1% agarose gel electrophoresis to check the quality of the sample.
[0142] (2) Take 1 μl of qualified DNA sample, and use ...
Embodiment 3
[0158] A kit and method for determining the genotype of the ABCC4 gene polymorphism site rs3742106 based on restriction fragment length polymorphism analysis.
[0159] 1. The composition of the kit:
[0160] 1. Primers for amplifying the polymorphic site rs3742106 of the ABCC4 gene:
[0161] SEQ ID No.2: 5'-GTTAGAGCTGGAGATCCTTG-3'
[0162] SEQ ID No.3: 5'-AGTCCGTTCCGAAGGCAATT-3'
[0163] 2. PCR amplification enzyme and corresponding buffer;
[0164] 3. dNTPs;
[0165] 4. Restriction endonuclease and its reaction buffer.
[0166] 2. Genotype detection
[0167] 1. Collect specimens
[0168] 1ml of whole blood was drawn from the patient's peripheral vein and placed in a 0.5M EDTA anticoagulant tube.
[0169] 2. Genomic DNA extraction
[0170] Use the Genomic DNA Extraction Kit to extract DNA according to the instructions.
[0171] 3. Genotype detection
[0172] (1) Take 1 μl of DNA sample, and use 1% agarose gel electrophoresis to check the quality of the sample.
[01...
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