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Real-time quantitative PCR

MZ Mengqi Zhang
BL Beifang Li
HL Haiyan Liao
ZC Zuhua Chen
WH Wenwen Huang
JY Jing Yang
SG Sai Ge
ZL Zhongwu Li
LS Lin Shen
CZ Cheng Zhang
JG Jing Gao
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Primer 5.0 software was used to design the primers. c-Myc forward primer: 5′-TCCTGTACCTCGTCCGATTC-3′, c-Myc reverse primer: 5′-GGTTTGCCTCTTCTCCACAG-3′; TCF7 forward primer: 5′-AGCACCAAGAATCCACCACA-3′, TCF7 reverse primer: 5′-GATTCCCACCACTTGGAGCA-3′; β-actin forward primer: 5′-GAGCTACGAGCTGCCTGACG-3′, β-actin reverse primer: 5′-CCTAGAAGCATTTGCGGTGG-3′. The extracted RNAs from 176P and 176R tissues were reversed to cDNAs followed by evaluating the expression level of target gene using TranStart Top Green qPCR SuperMix kit (TransGen Biotech, Beijing, China) based on the ABI 7500 real-time PCR system (Applied Biosystems). The expression of target gene was evaluated by the comparative Ct (ΔΔCT) method as the following formula.

(tg: target gene, rg: reference gene)

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