New application of KCNK2 gene
A gene and gene chip technology, which is applied in the fields of medical preparations containing active ingredients, determination/inspection of microorganisms, biochemical equipment and methods, etc. Difficulty in changing the organizational structure and other problems, to achieve the effect of simplifying the quantitative detection process, strong selectivity, and clear results
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Embodiment 1
[0031] The collection of embodiment 1 case
[0032] Samples from 5 cases of papillary thyroid carcinoma and 8 healthy controls were collected. The case samples were patients with papillary thyroid carcinoma confirmed by intraoperative frozen rapid pathology and postoperative paraffin slow pathology, which were confirmed by eight thyroid function tests and thyroid color Doppler ultrasound examination.
Embodiment 2
[0033] Example 2 High-throughput sequencing and analysis
[0034] use Reagent extracts sample RNA, and after RNA extraction, agarose gel electrophoresis is used to detect the extraction of RNA samples by NanoDrop1000 spectrophotometer. The sample requirements for RNA-seq sequencing: OD260 / OD280 is 1.8-2.2.
[0035] The HiSeq 2500 high-throughput sequencing platform of Illumina was used for high-throughput transcriptome deep sequencing, and the Fast-QC software was used to evaluate the quality of the sequencing data after sequencing, including the quality value distribution of bases, the position distribution of quality values, GC content, PCR duplication content, kmer frequency, etc. In the analysis of differential gene expression, according to the obtained FPKM value, the internationally recognized algorithm EBSeq was used for differential screening. Among them, when filtering, LOG 2 FC>1 or <-1, FDR<0.05. In order to better understand the functions of differentially expr...
Embodiment 3
[0036] Example 3 KCNK2 gene expression in papillary thyroid carcinoma group and control group
[0037] 1. Materials and methods
[0038] 1. Materials
[0039] During the period from May 2013 to November 2015, 45 cases of papillary thyroid carcinoma tissues and 18 cases of healthy control tissues were selected and grouped and numbered.
[0040] 2. Method
[0041] 2.1 Extraction of total RNA in papillary thyroid carcinoma group and control group
[0042] use Reagent extracts sample RNA, and the operation refers to the instructions, as follows:
[0043] After collecting the samples, freeze them in liquid nitrogen. After taking them out, put the tissue into a pre-cooled mortar for grinding. After the tissue sample is powdered:
[0044] ① Add Trizol and let stand at room temperature for 5 minutes;
[0045] ② Add 0.2ml chloroform to every ml Trizol, vibrate the centrifuge tube vigorously, mix well, and place it at room temperature for 5-10min;
[0046] ③ After high-speed cen...
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