Application of fingerprint spectrum consisting of microRNAs in diagnosis and treatment of human ovarian cancer
A technology for ovarian cancer and ovarian cells, applied in the field of small RNA (miRNA), can solve the problems of difficult diagnosis and effective treatment of ovarian cancer
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[0124] The preparation of the miRNA chip can adopt conventional manufacturing methods of biochips known in the art. For example, if the solid phase carrier is a modified glass slide or silicon wafer, and the 5' end of the probe contains amino-modified poly dT strings, the oligonucleotide probe can be formulated into a solution, and then spotted with a spotting instrument. The miRNA chip of the present invention can be obtained by arranging in a predetermined sequence or array on a modified glass slide or a silicon chip, and then fixing it by standing overnight. If the nucleic acid does not contain amino modification, its preparation method can also refer to: "Gene Diagnosis Technology-Non-radioactive Operation Manual" edited by Wang Shenwu; J.L.erisi, V.R.Iyer, P.O.BROWN. Exploring the metabolic and genetic control of gene expression on a genomic scale.Science,1997;278:680 and Ma Liren, edited by Jiang Zhonghua. Biochip. Beijing: Chemical Industry Press, 2000, 1-130.
[0125]...
Embodiment 1
[0148] Sample collection and information analysis
[0149] The samples were approved by the Ethics Committee of Shanghai Zhongshan Hospital, and were obtained in accordance with the collection method of the Shanghai Zhongshan Hospital tissue bank under the condition that the patients signed the informed consent. A total of 63 tissue samples were removed from the patients and stored at -80°C, and the sample information is shown in Table 1. Tissues used for biostatistical analysis included 14 normal ovarian tissue samples and 45 tumor samples, including 39 epithelial ovarian cancer samples and 6 epithelial borderline ovarian cancer tissue samples.
[0150] Table 1 Sample information
[0151]
Embodiment 2
[0153] Total RNA Extraction
[0154] Fragments less than 50 mg were cut from frozen or fresh animal tissues, weighed, and the remaining tissues were quick-frozen in liquid nitrogen.
[0155]After the excised fragments were homogenized, total RNA was extracted according to the instructions of miRNeasy MiniKit (Qiagen, 217004).
[0156] The mass was detected by electrophoresis, and the OD was determined by UV spectrophotometry 260nm and OD 280nm , to calculate the RNA concentration. Store at -80°C.
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