SNP marker related to auxiliary diagnosis of lung cancer of non-smoking women and applications thereof
An auxiliary diagnosis, non-smoking technology, applied in the field of SNP markers
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Embodiment 1
[0086] Example 1 The collection of samples and the arrangement of sample data
[0087] The inventor collected a large number of blood samples from non-smoking female lung cancer patients from April 2005 to December 2011 from the First Affiliated Hospital of Nanjing Medical University and Jiangsu Cancer Hospital. After sorting out the sample data, the inventor selected 1822 experimental samples of genome-wide microarray scanning and single SNP Sequenom MassARRAY genotyping of samples meeting the following criteria:
[0088] 1. Non-smoking female lung cancer patients diagnosed by histopathology;
[0089] 2. Non-smoking female healthy controls matched with the age of the case;
[0090] The demographic data and clinical data of these samples were collected systematically.
Embodiment 2
[0091] Example 2 Whole Genome Scanning of SNPs in Peripheral Blood DNA
[0092] Among the above-mentioned eligible 228 non-smoking female lung cancer patients and 568 non-smoking female healthy controls, the two groups were age-matched. The two groups of people were detected by Affymetrix6.0 chip to obtain relevant results. The specific steps are:
[0093] 1. Add hemolysis reagent (i.e. lysate, 40 parts) to the peripheral blood stored in the 2ml cryopreservation tube. Dilute the TrisHcl solution to 2000ml, the same below), turn it upside down and mix it completely.
[0094] 2. Removal of red blood cells: Fill the 5ml centrifuge tube to 4ml with hemolysis reagent, mix by inverting, centrifuge at 4000rpm for 10 minutes, and discard the supernatant. Add 4ml of hemolysis reagent to the precipitate, invert and wash again, centrifuge at 4000rpm for 10 minutes, and discard the supernatant.
[0095] 3. Extract DNA: Add 1ml extract solution to the precipitate (each 300ml contains 1...
Embodiment 3
[0105] Example 3 Sequenom MassARRAY genotyping of a single SNP
[0106] The SNPs found to be associated with the incidence of lung cancer in non-smoking women were detected by the above genome-wide scan on another 502 non-smoking female lung cancer cases and 524 non-smoking female healthy controls on the Sequenom MassARRAY genotyping platform. The specific steps are as follows:
[0107] 1. Add the hemolysis reagent to the peripheral blood stored in the 2ml cryopreservation tube, mix it upside down and transfer it completely.
[0108] 2. Removal of red blood cells: Fill the 5ml centrifuge tube to 4ml with hemolysis reagent, mix by inverting, centrifuge at 4000rpm for 10 minutes, and discard the supernatant. Add 4ml of hemolysis reagent to the precipitate, invert and wash again, centrifuge at 4000rpm for 10 minutes, and discard the supernatant.
[0109] 3. Extract DNA: Add 1ml of extract solution and 8μl of proteinase K to the precipitate, shake and mix well on the shaker, and ...
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