Application of SPAG7 (Sperm Associated Antigen) gene in preparing senile dementia diagnosis kit
A kind of Alzheimer's disease and gene technology, applied in the field of biomedicine, can solve the problems that need to be further verified, leave sequelae, and difficult to obtain the process, and achieve the effect of simplifying the quantitative detection process, strong selectivity, and good repeatability.
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Embodiment 1
[0029] Example 1 High-throughput sequencing and analysis
[0030] The samples came from Peking Union Medical College Hospital, and the informed consent of the subjects was obtained. Collect peripheral blood samples from 15 cases of senile dementia patients and 9 cases of healthy control peripheral blood samples for RNA extraction. After RNA extraction, agarose gel electrophoresis can be used to preliminarily determine whether the quality of the extracted RNA samples is qualified or not. for further transcriptome analysis. Then, the extraction of RNA samples was detected by NanoDrop1000 spectrophotometer, and the sample requirements for RNA-seq sequencing: OD260 / OD280 was 1.8-2.2.
[0031] The sequencing platform is the HiSeq 2500 high-throughput sequencing platform of Illumina, which performs high-throughput transcriptome deep sequencing. After sequencing, we use Fast-QC (http: / / www.bioinformatics.babraham.ac.uk / projects / fastqc / ) The software evaluates the quality of sequenc...
Embodiment 2
[0032] Example 2 SPAG7 gene expression in peripheral blood of senile dementia patients and healthy people
[0033] 1. Materials and methods
[0034] 1. Materials
[0035] The peripheral blood of 95 cases of senile dementia patients and 31 cases of healthy people were collected, grouped and numbered. 2. Method
[0036] 2.1 Extraction of total RNA from peripheral blood of senile dementia patients and peripheral blood of healthy people
[0037] use Reagent extracts sample RNA, and the experimental operation is carried out according to the product manual. For specific operations, see the manual.
[0038] Judgment criteria for RNA quality: the OD260 / OD280 value of the RNA sample is between 1.7 and 2.2; the electrophoretic pattern of total RNA has clear 28S and 18S bands; difference.
[0039] 2.2 Synthesis of cDNA by reverse transcription
[0040] use III Reverse Transcriptase (invitrogen, Cat. No. 18080-044) was used for cDNA reverse transcription, and the experimental op...
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