LNCaP cells were seeded to either 6 cm or 10 cm dishes at 800,000 or 2,200,000 cells/dish respectively. Cells were allowed to attach for 24 h and then subsequently treated with either enzalutamide (5 μM) or DMSO for specified time period. Cells were then harvested with Trizol reagent (Ambion) and RNA integrity was verified via agarose gel electrophoresis. Promega High Capacity cDNA Reverse Transcription Kit (Promega) was utilized following manufacturer instructions and as described previously [36] (link), [37] (link), [38] (link). RT-qPCR was performed with FastStart Universal SYBR Green Master Mix (Thermo Fisher Scientific) and detected on a QuantStudio 6 Flex with QuantStudio Real-Time PCR control software (Thermo Fisher Scientific). QuantStudio Design and Analysis software (Thermo Fisher Scientific) was used for data analysis. Technical triplicates were run for all samples, samples without detectable amplification were deemed undetected. Primer sets were validated via melt curve and agarose gel analysis of RT-qPCR product. AR primers were used as described previously [36] (link) and IVL primers were used as described previously [39] (link). All primer sequences utilized are described in Supplementary Table S1 .
Quantstudio real time pcr control software
Manufactured by Thermo Fisher Scientific
The QuantStudio Real-Time PCR control software is a software application that provides control and data analysis capabilities for QuantStudio real-time PCR instruments. The software enables users to set up, run, and analyze real-time PCR experiments.
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Lab products found in correlation
Faststart universal sybr green master mix, by Thermo Fisher Scientific (2 mentions)
Quantstudio 6 flex, by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Quantstudio design and analysis software, by Thermo Fisher Scientific (2 mentions)
High capacity cdna reverse transcription kit, by Promega (2 mentions)
2 protocols using quantstudio real time pcr control software
1
Enzalutamide Regulation of Gene Expression
2
Transcriptional Regulation of Androgen Receptor in Prostate Cancer
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LNCaP cells were seeded to either 6 cm or 10 cm dishes at 800,000 or 2,200,000 cells/dish respectively. Cells were allowed to attach for 24 h and then subsequently treated with either compound 17 (500 nM) or DMSO for 72 h. Cells were then harvested with Trizol reagent (Ambion) and RNA integrity was verified via agarose gel electrophoresis. Promega High Capacity cDNA Reverse Transcription Kit (Promega) was utilized following manufacturer instructions and as described previously 41, (link)71, (link)72 (link) . RT-qPCR was performed with FastStart Universal SYBR Green Master Mix (Thermo Fisher Scientific) and detected on a QuantStudio 6 Flex with QuantStudio Real-Time PCR control software (Thermo Fisher Scientific). QuantStudio Design and Analysis software (Thermo Fisher Scientific) was used for data analysis. Technical triplicates were run for all samples, samples without detectable amplification were deemed undetected. Primer sets were validated via melt curve and agarose gel analysis of RT-qPCR product. AR primers were used as described previousy 41 (link) and IVL primers were used as described previously. 40 All primer sequences utilized are described in Supplementary Information.
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