Application of LOC100129066 in preparation of diagnostic product for prostatic cancer
A technology for prostate cancer and products, applied in the application field of preparing prostate cancer diagnosis products, can solve the problems of not very stable diagnosis scheme, misdiagnosed disease, failure to achieve early diagnosis, etc., and achieve the effect of rapid and effective early detection
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Embodiment 1
[0031] Example 1 High-throughput sequencing screening of differentially expressed genes
[0032] 1. Sampling
[0033] Select 10 cases of fresh prostate cancer tissues and corresponding paracancerous tissues, samples from prostate cancer patients who underwent radical prostatectomy and bilateral pelvic lymph node dissection from October 2012 to December 2015 in Peking Union Medical College Hospital. Under the guidance of the pathologist, the prostate cancer and paracancerous tissues were immediately taken and put into liquid nitrogen, and stored in a -80°C low-temperature refrigerator after serial numbering.
[0034] 2. Total RNA extraction from tissue samples
[0035] use Reagent (invitrogen, Cat. No. 15596-018) was used to extract sample RNA, and the experimental operation was carried out according to the product manual. The specific operation was as follows:
[0036] Collect the samples and freeze them in liquid nitrogen. After taking them out, put the tissue samples int...
Embodiment 2
[0047] Example 2 RT-PCR verification of the expression of LOC100129066 in prostate cancer tissue
[0048] 1. Materials
[0049] 10 prostate cancer patients were selected, and the samples were collected from the prostate cancer patients who underwent radical prostatectomy and bilateral pelvic lymph node dissection from October 2012 to December 2015 in Peking Union Medical College Hospital. Immediately take prostate cancer and paracancerous tissues and put them into liquid nitrogen, and store them in a -80°C low-temperature refrigerator after serial numbering.
[0050] 2. Method
[0051] 2.1 Extract the total RNA from the tissue samples of the subjects, the same as the extraction method in Example 1.
[0052] 2.2 Synthesis of cDNA by reverse transcription
[0053] use III Reverse Transcriptase (invitrogen, Cat. No. 18080-044) was used for cDNA reverse transcription, and the experimental operation was carried out according to the product manual. The specific operation was as...
Embodiment 3
[0075] Example 3 Application of Gene Chip Diagnosis of Prostate Cancer
[0076] 1, the acquisition of material, with embodiment 2.
[0077] 2, the extraction of total RNA, with the method for embodiment 1.
[0078] 3. Gene chip verification
[0079]After the total RNA was linearized and amplified, the cy3-UTP labeled and fluorescently labeled cRNAs were purified using the RNEASY Mini Kit, and the labeled cRNAs were fragmented with Amhion’s RNA Fragmentation Reagents. Human whole gene expression profile chips (4x 44K genes) from Agilent Company of the United States were used, hybridized in a chip hybridization oven at 65°C for 17 hours, then eluted, stained, and finally scanned with an Agilent DNA MicroarrayScanner scanner.
[0080] After the hybridized chip reads the data points by the chip scanner, the data is imported into the analysis software, and the genes whose absolute value of the natural logarithm of the ratio of the two groups is greater than 2.0 or less than 0.5 a...
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