Primers and probes for analysis were selected using Beacon Designer 7.0 (Premier Biosoft International, USA) and corresponding sequences from the NCBI database57 . The specificities of primers and probes were checked using Primer358 , Primer-BLAST59 .
The relative levels (R) of transcripts in the test groups were calculated as R = 2−ΔΔCt60 (link). That in the control group was set as 1. Statistical data processing was performed using Statistica for Windows 8.061 and Microsoft Excel 2016 (Microsoft, USA). The relative levels of gene expression were assessed using the Mann–Whitney nonparametric U test. Receiver operator characteristic (ROC) curves, and qPCR efficiency and specificity were obtained using the Real Statistics Resource Pack of MS Excel 2016 (Microsoft, USA). Gene interaction networks were built using Pathway Studio® version 12.1.0.9 (Elsevier, Netherlands)62 (link),63 (link).
The relative levels (R) of transcripts in the test groups were calculated as R = 2−ΔΔCt60 (link). That in the control group was set as 1. Statistical data processing was performed using Statistica for Windows 8.061 and Microsoft Excel 2016 (Microsoft, USA). The relative levels of gene expression were assessed using the Mann–Whitney nonparametric U test. Receiver operator characteristic (ROC) curves, and qPCR efficiency and specificity were obtained using the Real Statistics Resource Pack of MS Excel 2016 (Microsoft, USA). Gene interaction networks were built using Pathway Studio® version 12.1.0.9 (Elsevier, Netherlands)62 (link),63 (link).
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