Genome extraction kit

In the experiment, 33 OSCC patients treated in the Affiliated Hospital of Taishan Medical University (Taian, China) from January 2016 to January 2018 were selected as the experimental group, including 18 males and 15 females, aged 45 years on average, while 33 healthy subjects were selected as the control group, including 17 males and 16 females, aged 45 years on average. The experimental scheme was discussed and approved by the Academic Committee, and agreed by the family members.
This study was approved by the Ethics Committee of Affiliated Hospital of Taishan Medical University. Patients who participated in this research had complete clinical data. The signed informed consents were obtained from the patients or the guardians.
The following experimental reagents were used: Fluorescence quantitative polymerase chain reaction (PCR) reagent and ribonucleic acid (RNA) extraction reagent were purchased from Takara and animal cell protein extraction kit was purchased from Thermo Fisher Scientific, Inc. Streptomycin-peroxidase (S-P) kit and corresponding antibodies were purchased from Thermo Fisher Scientific, Inc. Genome extraction kit was purchased from AXYGEN and other reagents and consumables were purchased from Sangon Biotech Co., Ltd.

The OSCC cell lines including SCC-9, SCC-25, HN4, Tca-811 and hNOK cells were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Human normal oral keratinocyte cell line hNOK cells were used as the negative control. For cell lines, we extracted DNA from various cells (SCC 9, SCC 25, HN4, Tca 811 and hNOK) using the Genome extraction kit of Axygen, amplified by the 21-STR amplification scheme. And then we detected STR spot and sex gene Amelogenin on the ABI 7900HT Fast Real-Time PCR System (Supplementary Table 2–6). No multiple alleles were found in these cell lines in this test. All cells were cultured in DMEM medium supplemented with 10% FBS and 100 μg/mL penicillin/streptomycin at 37°C with 5% CO₂.