Breast cancer recurrent risk assessment 21 gene detection primer and application thereof
A technology of risk assessment and genetic detection, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of high recurrence and transfer rates, shorten the time of detection experiments, improve efficiency, expand conditionally consistent effect
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Embodiment 1
[0050] Example 1: Detecting the nucleic acid sequences of the 21 genes detected in breast cancer recurrence risk assessment.
[0051] 21 pairs of specific primers were entrusted to Yingwei Jieji (Shanghai) Trading Co., Ltd. to synthesize:
[0052] SEQ ID NO: 1 5'-CGGACTTTGGGTGCGACTT-3'
[0053] SEQ ID NO: 2 5'-TTACAACTCTTCCACTGGGACGAT-3'
[0054] SEQ ID NO: 3 5'-GCTAGAGGCATCATGGACCGA-3'
[0055] SEQ ID NO: 4 5'-GAAGGACACAAGACCCGCTGA-3'
[0056] SEQ ID NO: 5 5'-AGAACTGGCCCTTCTTGGAGG-3'
[0057] SEQ ID NO: 6 5'-TCTATGGGGTCGTCATCTGGC-3'
[0058] SEQ ID NO: 7 5'-TTCAGGTTGTTGCAGGAGAC-3'
[0059] SEQ ID NO: 8 5'-CATCTTCTTGGGCACACAAT-3'
[0060] SEQ ID NO: 9 5'-GCCGAGATCGCCAAGATG-3'
[0061] SEQ ID NO: 10 5'-CTTTTGATGGTAGAGTTCCAGTGATTC-3'
[0062] SEQ ID NO: 11 5'-CCATCTGCATCCATCTTGTT-3'
[0063] SEQ ID NO: 12 5'-GGCCACCAGGGTATTATCTG-3'
[0064] SEQ ID NO: 13 5'-CGGTGTGAGAAGTGCAGCAA-3'
[0065] SEQ ID NO: 14 5'-CCTCTCGCAAGTGCTCCAT-3'
[0066] SEQ ID NO: 15 5'-TGTCCAGCCACC...
Embodiment 2
[0094] Example 2: 21-gene detection method for breast cancer recurrence risk assessment.
[0095] Instrument: Roche 480 fluorescent quantitative PCR detector, BECKMAN 22R desktop micro-refrigerated centrifuge, Eppendorf 5810R desktop refrigerated centrifuge, Taicang Hualida Laboratory Equipment Company WH-866 vortex oscillator.
[0096] (1) Paraffin tissue RNA extraction reagent
[0097] Paraffin tissue RNA was extracted according to the instructions of the paraffin tissue RNA extraction kit produced by Qiagen Biological Company, and tissue RNA was obtained for future use.
[0098] (2) reverse transcription, the reagents were purchased from THERMO company
[0099] The reaction system is a 20 μL system, including 11 μL template RNA, 1 μL RT enzyme, 1 μL RI enzyme, 2 μL dNTP, 4 μL 5×buffer, and 1 μL random primer; reaction conditions: 42°C for 30 min, 95°C for 5 min.
[0100] Prepare 3 tubes for one specimen.
[0101] (3) Fluorescent quantitative PCR, the reagents were pu...
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