A quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) was performed as previously described [13 (link)]. Briefly, RNA was isolated from cell cultures using TRIzol® reagent according to the supplier’s instructions (Thermo Fisher Scientific, Breda, The Netherlands). RNA quality and quantity were determined using a NanoDrop 2000c UV–vis spectrophotometer (Thermo Fisher Scientific). The cDNA was synthesized from 2.5 µg RNA using random nanomers and M-MLV reverse transcriptase (Invitrogen, Carlsbad CA, USA). Taqman primers and probes were designed using Primer Express 3.0.1 and are shown in Supplementary Table S1 . All target genes were amplified using the Q-PCR core kit master mix (Eurogentec, Liège, Belgium) on a 7900HT Fast Q-PCR system (Thermo Fisher Scientific). SDSV2.4.1 (Thermo Fisher Scientific) software was used to analyze the data. Expression of genes is presented in 2-delta CT and normalized to 18S.
Q pcr core kit master mix
Manufactured by Eurogentec
Sourced in Belgium
The Q-PCR core kit master mix is a ready-to-use solution designed for quantitative real-time PCR (qPCR) experiments. It contains all the necessary components, including DNA polymerase, dNTPs, and buffer, to perform qPCR reactions.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (8 mentions)
M mlv reverse transcriptase, by Thermo Fisher Scientific (5 mentions)
Nanodrop 2000c uv vis spectrophotometer, by Thermo Fisher Scientific (4 mentions)
Sds v2, by Thermo Fisher Scientific (3 mentions)
7900ht fast real time pcr system, by Thermo Fisher Scientific (3 mentions)
7900ht fast qpcr system, by Thermo Fisher Scientific (1 mentions)
Primer express 3, by Thermo Fisher Scientific (1 mentions)
4 protocols using q pcr core kit master mix
1
Quantitative Real-Time RT-PCR for Gene Expression
2
Quantitative RT-PCR for Gene Expression
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Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) was performed as previously described [10 (link)]. Briefly, RNA was isolated from cell cultures using TRIzol® reagent according to the supplier’s instruction (Thermo Fisher,). RNA quality and quantity were determined using a Nanodrop 2000c UV-vis spectrophotometer (Thermo Fisher). cDNA was synthesized from 2.5 µg RNA using random nanomers and M-MLV reverse transcriptase (Invitrogen). Taqman primers and probes were designed using Primer Express 3.0.1 and are shown in Supplementary Table S1 . All target genes were amplified using the Q-PCR core kit master mix (Eurogentec, Seraing, Belgium) on a 7900HT Fast Real-Time PCR system (Thermo Fisher). SDSV2.4.1 (Thermo Fisher) software was used to analyze the data. Expression of genes is presented in 2-delta CT and normalized to 18S.
3
Quantitative Real-Time RT-PCR for Gene Expression
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Quantitative real-time reverse transcription polymerase chain reaction was performed as previously described.58 (link) Briefly, total RNA was isolated from tissue samples using TRIzol reagent according to supplier’s instructions (Thermo Fisher Scientific). RNA quality and quantity were determined using a Nanodrop 2000c UV-vis spectrophotometer (Thermo Fisher Scientific). Complementary DNA was synthesized from 2.5 μg of RNA by using random nonamers and M-MLV reverse transcriptase (Thermo Fisher Scientific). TaqMan primers and probes were designed using Primer Express 3.0.1 (Thermo Fisher Scientific) and are shown in Supplementary Table 2 . All target genes were amplified using the Q-PCR core kit master mix (Eurogentec, Maastricht, the Netherlands) on a 7900HT Fast Real-Time PCR system (Thermo Fisher Scientific). SDSV2.4.1 (Thermo Fisher Scientific) was used to analyze the data. Expression of genes is presented in 2-ΔCT and normalized to 36B4.
4
Quantitative RT-PCR Analysis of Gene Expression
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Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) was performed as previously described [25] (link). Shortly, total RNA was isolated from tissue samples using TRIzol® reagent according to supplier's instruction (ThermoFisher Scientific, The Netherlands). RNA quality and quantity were determined using a Nanodrop 2000c UV-vis spectrophotometer (ThermoFisher Scientific). cDNA was synthesized from 2.5 μg RNA using random nonamers and M-MLV reverse transcriptase (Invitrogen, USA). Taqman primers and probes were designed using Primer Express 3.0.1 and are shown in Supplementary Table S1. All target genes were amplified using the Q-PCR core kit master mix (Eurogentec, The Netherlands) on a 7900HT Fast Real-Time PCR system (Applied Biosystems Europe, The Netherlands). SDSV2.4.1 (Applied Biosystems Europe, The Netherlands) software was used to analyze the data. Expression of genes was normalized to 18S and presented as relative levels compared to a control condition set to 1.
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