Breast cancer molecular marker miR-30c-1-3p
A molecular marker and breast cancer technology, applied in the field of tumor diagnosis, can solve the problems of miRNA marker accuracy, insufficient sensitivity, inability to display the whole picture of lesions, and inability to provide qualitative diagnosis, etc., to achieve easy mass screening and high specificity , The effect of convenient material collection
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[0017] 1. Extraction of total RNA from serum
[0018] Take 2ml of blood from 5 breast cancer patients before operation and 7 days after operation, and take 2ml of blood from 5 normal people as a normal control. in a centrifuge tube.
[0019] Total RNA was extracted from serum using RNA extraction kit (Beijing Kuangbo Biotechnology Co., Ltd.), and 1 μl (20 nM) of the sequence 5'-CAACCUCCUAGAAAGA-3' (SEQ ID NO: 2) was added to each 250 μl of serum. 1 (synthesized by Shanghai Sangon Bioengineering Technology Co., Ltd.) to monitor the quality of RNA extraction from serum. The concentration of extracted total RNA was measured using Thermo NanoDrop2000c.
[0020] 2. Three-step method for quantitative detection of miRNA in serum
[0021] (1) Add polyA tail:
[0022] i. Prepare the polyA-tailed reaction solution in an RNase-free PCR tube (Axygen, 200 μl), with a volume of 20 μl. Add 1 μl (20nM) External Control-2 (synthesized by Shanghai Sangon Bioengineering Technology Co., Ltd....
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