RNA was extracted with Trizol (Thermo Fisher Scientific, Waltham, MA, USA) and reversely transcribed to cDNA using a RevertAid First Strand cDNA Synthesis kit (Fermentas, Vilnius, Lithuania). The cDNA was subjected to either the conventional RT-PCR analysis using Taq PCR Mastermix (Tiangen biotechnology, Beijing, China) on an Applied Biosystems Veriti 96 well Thermal Cycler PCR detection system (Thermo Fisher Scientific) or the quantitative RT-PCR analysis using iQ SYBR Green supermix (BioRad Laboratories, Hercules, CA, USA) on an iQ5 qRT-PCR detection system (Bio-Rad Laboratories). The primer sequences synthesized by the Beijing AuGCT DNA-SYN Biotechnology Company (Beijing, China) were summarized in Table S1. All experiments were performed in triplicate. The expression level of each gene was normalized to the level of housekeeping gene, Gapdh.
Applied biosystems veriti 96 well thermal cycler pcr detection system
Manufactured by Thermo Fisher Scientific
Sourced in China
The Applied Biosystems Veriti 96 Well Thermal Cycler is a PCR detection system designed for amplifying DNA samples. It features a 96-well reaction block and can precisely control the temperature of each sample well during the thermal cycling process.
Automatically generated - may contain errors
Lab products found in correlation
Iq sybr green supermix, by Bio-Rad (2 mentions)
Taq pcr mastermix, by Tiangen Biotech (2 mentions)
Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Iq5 qrt pcr detection system, by Bio-Rad (1 mentions)
Quantstudio five real time pcr system, by Thermo Fisher Scientific (1 mentions)
2 protocols using applied biosystems veriti 96 well thermal cycler pcr detection system
1
Gene Expression Analysis by RT-PCR
2
RT-PCR and qRT-PCR Analysis of Gene Expression
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The cDNA was subjected to either the conventional RT-PCR analysis using Taq PCR Mastermix (Tiangen biotechnology, Beijing, China) on an Applied Biosystems Veriti 96 well Thermal Cycler PCR detection system (Thermo Fisher Scientific) or the quantitative RT-PCR analysis using iQ SYBR Green supermix (BioRad Laboratories, Hercules, CA, USA) on a QuantStudio five Real-Time PCR System (Thermo Fisher Scientific). Relative quantification for gene expression was calculated using the 2−ΔΔCT method, which was normalized to the internal reference, β-actin. The primer sequences were summarized in Table S1 .
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