Reverse Transcription (RT) was performed with 1 – 2 μg total RNA and using 3.2 μg random hexamer primers (ThermoFisher Scientific, Cat#SO142) and Transcriptor Reverse Transcriptase (Roche, Cat#03531287001) following manufacturer’s instruction. For qPCR, the cDNA samples were amplified using LightCycler® 480 SYBR® Green 2x PCR Master Mix I (Roche, Cat#04887352001) and 0.6 μM of forward and reverse primer respectively. The primer pairs used for qPCR are listed in Table S2 . The qPCR was conducted using a LightCycler® 480 (Roche). The obtained threshold-values were used to calculate the relative gene expression using the 2−ΔΔCT method and considering the individual primer pair efficiencies (Pfaffl, 2001 (link)).
Lightcycler 480 sybr green 2x pcr master mix 1
Manufactured by Roche
The LightCycler® 480 SYBR® Green 2x PCR Master Mix I is a ready-to-use reaction mix for real-time PCR analysis. It contains a DNA polymerase, SYBR® Green I dye, and necessary reagents for amplification and detection of DNA sequences.
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2 protocols using lightcycler 480 sybr green 2x pcr master mix 1
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Quantitative Real-Time PCR Analysis
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Quantitative Real-Time PCR Analysis
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Reverse Transcription (RT) was performed with 1 – 2 μg total RNA and using 3.2 μg random hexamer primers (ThermoFisher Scientific, Cat#SO142) and Transcriptor Reverse Transcriptase (Roche, Cat#03531287001) following manufacturer’s instruction. For qPCR, the cDNA samples were amplified using LightCycler® 480 SYBR® Green 2x PCR Master Mix I (Roche, Cat#04887352001) and 0.6 μM of forward and reverse primer respectively. The primer pairs used for qPCR are listed in Table S2 . The qPCR was conducted using a LightCycler® 480 (Roche). The obtained threshold-values were used to calculate the relative gene expression using the 2−ΔΔCT method and considering the individual primer pair efficiencies (Pfaffl, 2001 (link)).
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