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ChIP-quantitative PCR (ChIP-qPCR) analysis

JR Junxiao Ren
WT Weihua Tian
KJ Keren Jiang
ZW Zhang Wang
DW Dandan Wang
ZL Zhuanjian Li
FY Fengbin Yan
YW Yanbin Wang
YT Yadong Tian
KO Kepeng Ou
HW Hongjun Wang
XK Xiangtao Kang
HL Hong Li
XL Xiaojun Liu
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The immunoprecipitated DNA fragments were also used to verify the interaction of ERα and selected binding sites by ChIP-qPCR. Meanwhile, 20% of starting chromatin without chromatin immunoprecipitation served as input to represent the unselected DNA content. The fold enrichment method was chosen to normalize the ChIP-qPCR data: Fold enrichment = log2−ΔΔCt, ΔCt = Ct (IP) − Ct (Input) − log25, ΔΔCt = ΔCt − ΔCt (IgG). Gene-specific primers for the putative binding-site regions were designed using the NCBI Primer-BLAST tool and synthesized by Sangon Biotech (Shanghai, China). All ChIP-qPCR primer sequences used are listed in Table S7.

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