Comprehensive RNA Analysis Protocol

Total RNA from cells was extracted using TRIzol reagent (Invitrogen, United States) as previously described [20 (link)]. The complementary DNA was synthesized with random hexamer primer using Verso cDNA Synthesis Kit (Thermo Fisher Scientific, United States). Quantitative real-time PCR (qRT-PCR) was carried out on ABI StepOne Plus real time PCR system (Applied Biosystems, United States) using SYBR mix kit (Thermo Fisher Scientific, United States) as previously described [21 (link)]. Primers for the 80 NEPC related chimeric RNAs were listed in Additional file 8: Table S1. Touch-down PCR (TD-PCR) was carried out using Platinum Taq High Fidelity Kit Invitrogen, United States. Primers for TMPRSS2-ERG (e1e4) and TMPRSS2-ERG (e2e4) were listed in Additional file 12: Table S5.

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Wang Q., Chen J., Singh S., Xie Z., Qin F., Shi X., Cornelison R., Li H, & Huang H. (2022). Profile of chimeric RNAs and TMPRSS2-ERG e2e4 isoform in neuroendocrine prostate cancer. Cell & Bioscience, 12, 153.

Publication 2022

TRIzol reagent Verso cDNA Synthesis Kit ABI StepOne Plus real time PCR system Platinum Taq High Fidelity Kit

Cdna Cells Chimeric Platinum Primers Quantitative real time pcr Rnas Synthesis Tmprss2 Touch Trizol

Corresponding Organization :

Other organizations : Sun Yat-sen Memorial Hospital, Sun Yat-sen University, University of Virginia

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression levels of 80 NEPC related chimeric RNAs
Expression levels of TMPRSS2-ERG (e1e4) and TMPRSS2-ERG (e2e4)

control variables

None explicitly mentioned

controls

No positive or negative controls were explicitly mentioned in the input text.

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