Quantitative Analysis of TopBP1 Expression

Trizol reagent (TaKaRa Biotechnology, Dalian, China) was used for extracting total RNA from cells and total RNA was reverse transcribed with a PrimerScript RT-PCR kit (Takara Biotechnology, Dalian, China) [31 (link)]. The Taqman TM RT-PCR system (Applied Biosystems, Carlsbad, USA) was used to conduct quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) according to the manufacturer’s protocol. The primers used are follows: TopBP1 forward: 5’- TTCAGCAACTCACAGTTAAGCA-3’ and reverse: 5’ GGCACACTCATACTTCTGACC-3’. PCR was performed in the presence of SYBR green (Sigma-Aldrich, St. Louis, USA) using 15 ng of total RNA with an ABI PRISM 7700 Sequence Detection System (Applied Biosystems).