Preparation of cDNA was performed as described, using both oligo (dT) and random hexamers (15 (link)). For strand-specific RT-qPCR, the indicated reverse transcription primers were used in the RT reaction, followed by inactivation of the RT enzyme. The resulting cDNA was used in qPCR reactions with Biorad SYBR Green Kit (Biorad) on a Mastercycler Realplex2 system (Eppendorf) and analyzed as described (15 (link)). Error bars represent the SEM from at least two technical repeats and two biological repeats per experiment. To determine the statistical significance of observed differences, p-values were calculated using Student’s t-test with p-values <0.05 considered significant.
Biorad sybr green kit
Manufactured by Bio-Rad
The Bio-Rad SYBR Green Kit is a reagent system designed for real-time quantitative PCR (qPCR) analysis. The kit contains SYBR Green I dye, which binds to double-stranded DNA and emits a fluorescent signal that is detected and measured by real-time PCR instruments. This allows for the quantification of target DNA sequences in a sample.
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Lab products found in correlation
2 protocols using biorad sybr green kit
1
Strand-Specific RT-qPCR Methodology
2
Quantitative PCR for Fecal Microbiome Analysis
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A Bio-Rad CFX100 sequence detection equipment and a Bio-Rad SYBR Green kit were used to conduct quantitative PCR (Bio-Rad). The specificity of the amplification products was confirmed by melting-point determination analysis. Total DNA was extracted from fecal samples of both cohorts adopting THSTI method (Bag et al. 201739 ). The copy counts of target genes in each sample were estimated by comparing the Ct values obtained from the standard curves with the control gene. The recA-specific primers recA-F/recA-R was employed as an internal control in each experiment. The results are given as the averages of qPCR data performed in triplicate. The fold change and p value derived with the t-test were then used to identify differentially expressed mRNA levels.
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