Gene detection composition and its application
A composition and gene technology, applied in the CCDC72 and ERC1 gene composition, the detection of postmenopausal osteoporosis and the clinical application of Liuwei Dihuang Pill, TMED2, the field of gene detection composition, can solve the unproven correlation, etc. problems, to achieve the effect of simplifying the quantitative detection process, high sensitivity, and clear results
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Embodiment 1
[0049] Example 1 High-throughput sequencing and analysis
[0050] The peripheral blood samples of 3 postmenopausal osteoporosis patients before and after taking Liuwei Dihuang Pills and 3 healthy controls were collected for RNA extraction. After RNA extraction, agarose gel electrophoresis was performed. Whether the quality of the RNA sample is qualified or not, whether it can be used 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.
[0051] 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 software to evaluate the quality of the sequencing data as a whole, including the quality value distribution of bases, quality value position distribution, GC content, PCR duplication content, kmer frequency, et...
Embodiment 2
[0052] Example 2 Peripheral blood TMED2, CCDC72 and ERC1 gene expression in postmenopausal osteoporosis patients before and after taking Liuwei Dihuang Pills
[0053]1. Materials and methods
[0054] 1. Materials
[0055] The consent of 22 patients with postmenopausal osteoporosis was collected, and the peripheral blood was collected at the initial stage of testing. After taking Liuwei Dihuang Pills for half a year, the peripheral blood was collected again, and the samples were grouped and numbered.
[0056] 2. Method
[0057] 2.1 Extraction of total RNA from peripheral blood of postmenopausal osteoporosis and healthy people
[0058] use Reagent was used to extract RNA from samples, and the experimental operation was performed according to the product instructions.
[0059] Judgment criteria for RNA quality: the OD260 / OD280 value of the RNA sample is between 1.8 and 2.2; the electrophoretic pattern of total RNA has clear 28S and 18S bands; difference.
[0060] 2.2 Synth...
Embodiment 3
[0083] Example 3 TMED2, CCDC72 and ERC1 gene expression in peripheral blood of postmenopausal osteoporosis patients and healthy people
[0084] 1. Materials
[0085] The peripheral blood of 32 postmenopausal osteoporosis patients and 29 healthy people were collected, grouped and numbered.
[0086] 2. Method
[0087] Concrete steps refer to embodiment 3.
[0088] 3. Results
[0089] Real-time quantitative PCR result display (see figure 2 ): The results of qRT-PCR amplification were stable, and the expression levels of TMED2, CCDC72 and ERC1 in the peripheral blood of postmenopausal osteoporosis were significantly different from those of healthy people. The former of the three genes was about 1.22% of the latter. times, 0.83 times and 1.39 times, the above results verified the results of high-throughput sequencing.
[0090] The present invention uses high-throughput sequencing to screen out postmenopausal osteoporosis-related genes TMED2, CCDC72 and ERC1, and further analy...
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