LYRM2 (LYR motif-containing 2) gene and new use of expression product thereof
A gene expression and gene technology, which is applied in the new application field of LYRM2 gene and its expression products in the detection of osteoporosis, can solve the problems of unclearness and lack of research results, and achieve clear results, strong selectivity and specificity. strong effect
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Embodiment 1
[0030] Example 1 High-throughput sequencing and analysis
[0031] Collect peripheral blood samples from 9 cases of osteoporosis patients and 6 cases of healthy people 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.
[0032] The sequencing platform is the HiSeq2500 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 / ) software Overall assessment of the quality of sequencing data, including base quality value distribution, quality value position distribution, GC content, PCRduplication content, kmer frequency...
Embodiment 2
[0033] Example 2 The expression of LYRM2 gene in the peripheral blood of patients with osteoporosis and the peripheral blood of healthy people
[0034] 1. Materials and methods
[0035] 1. Materials
[0036] The peripheral blood of 74 patients with osteoporosis and 46 healthy people were collected, grouped and numbered.
[0037] 2. Method
[0038] 2.1 Extraction of total RNA from peripheral blood of osteoporosis and healthy people
[0039] use Reagent was used to extract RNA from samples, and the experimental operation was performed according to the product instructions.
[0040] 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.
[0041] 2.2 Synthesis of cDNA by reverse transcription
[0042] use IIIReverseTranscriptase (invitrogen, Cat. No. 18080-044) was used for cDNA reverse transcription, and the experimental operation was carried out...
Embodiment 3
[0060] The cultivation of embodiment 3 cell lines MG-63
[0061] 1. Materials
[0062] The cell line MG-63 was purchased from Shanghai Cell Institute, Chinese Academy of Sciences.
[0063] 2. The main solution
[0064] Cell culture medium: DMEM medium + 10% standard fetal bovine serum.
[0065] 0.25% trypsin digestion solution: add 0.25g trypsin to 100m1 deionized water, filter and sterilize, and distribute for use.
[0066] 3. Cell passage
[0067] 1 Discard the original culture medium in the culture bottle full of cells, add 1m1 of 0.25% trypsin solution to cover the cell layer, sterilize the bottle mouth, and cover it;
[0068] 2 Observe the cell changes under an inverted microscope. As time goes by, the original adherent cells gradually tend to be round, the intercellular matrix shrinks, and the intercellular space increases. Discard the trypsin before floating and add 5ml The culture solution containing 10% fetal bovine serum terminates the digestion;
[0069] 3 Cel...
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