Total RNA was harvested by using the Rneasy Kit (Qiagen, Valencia, CA, USA). CDNA synthesis was done with Superscript II (40 U/l, Invitrogen, Groningen, The Netherlands). Real-time PCR was performed using the Biorad-iCycler IQ detector system and Biorad-iCycler IQ SYBR Green mix (Biorad, Munich, Germany). The PCR primers (Sigma Chemical Company, St. Louis, MO, USA) were: Shh 5'-CAGCGACTTCCTCACTTTCC-3' (forward) and 5'-GGAGCGGTTAGGGCTACTCT-3' (reverse); GAPDH: 5'-ACAGTCAGCCGCATCTTCTT-3' (forward) and 5'-ACGACCAAATCCGTTGACTC-3' (reverse). Fluorescent threshold values were measured in triplicate, and fold changes were calculated by the formula 2-(sample 1 ΔCt - sample 2 ΔCt), where ΔCt is the difference between the amplification fluorescence thresholds of the mRNA of interest and the GAPDH mRNA.
Biorad icycler iq detector system
Manufactured by Bio-Rad
Sourced in Germany
The Biorad-iCycler IQ detector system is a real-time PCR detection system designed for quantitative gene expression analysis. It features a sensitive optical detection system and a thermal cycler for precise temperature control during the amplification process. The core function of the Biorad-iCycler IQ detector system is to accurately measure and analyze fluorescent signals generated during the real-time PCR reaction.
Automatically generated - may contain errors
2 protocols using biorad icycler iq detector system
1
Quantitative RT-PCR Analysis of Shh Expression
2
Quantitative RT-PCR Analysis of Shh Expression
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