Yeast RNA was extracted using the YeaStar kit (Zymogen), RNA from Drosophila S2R cells was extracted using Trizol (Life Technologies), and RNA from human cells was extracted using the RNeasy PLUS mini kit (Qiagen). Human tissue RNA was obtained from Life Technologies (Human Brain Total RNA (AM7962), Human Heart Total RNA (AM7966) and Human Testes Total RNA (AM7972)). 500ng of RNA was used with the iScript cDNA synthesis Kit (BioRad), and 0.5μl of cDNA was used for each qPCR reaction, utilizing the KAPA SYBR® FAST Universal 2X qPCR Master Mix. The Drosophila qPCR reaction used iQ SYBR. qPCR primers are listed within the Supplementary Table 1 . qRT-PCR was run and analyzed on the CFX96 Real-Time PCR Detection System (BIORAD), with all target gene expression levels normalized to β-actin mRNA levels (human and M. musculus), FBA1 mRNA levels (S. cerevisiae) or RpL32 mRNA levels (D. melanogaster).
Kapa sybr fast universal 2x qpcr master mix
Manufactured by Bio-Rad
KAPA SYBR® FAST Universal 2X qPCR Master Mix is a ready-to-use solution for quantitative PCR (qPCR) reactions. It contains all the necessary components, including a high-performance DNA polymerase, for efficient and sensitive qPCR amplification.
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Lab products found in correlation
2 protocols using kapa sybr fast universal 2x qpcr master mix
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Quantitative Gene Expression Analysis
2
Quantitative Gene Expression Analysis
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Yeast RNA was extracted using the YeaStar kit (Zymogen), RNA from Drosophila S2R cells was extracted using Trizol (Life Technologies), and RNA from human cells was extracted using the RNeasy PLUS mini kit (Qiagen). Human tissue RNA was obtained from Life Technologies (Human Brain Total RNA (AM7962), Human Heart Total RNA (AM7966) and Human Testes Total RNA (AM7972)). 500ng of RNA was used with the iScript cDNA synthesis Kit (BioRad), and 0.5μl of cDNA was used for each qPCR reaction, utilizing the KAPA SYBR® FAST Universal 2X qPCR Master Mix. The Drosophila qPCR reaction used iQ SYBR. qPCR primers are listed within the Supplementary Table 1 . qRT-PCR was run and analyzed on the CFX96 Real-Time PCR Detection System (BIORAD), with all target gene expression levels normalized to β-actin mRNA levels (human and M. musculus), FBA1 mRNA levels (S. cerevisiae) or RpL32 mRNA levels (D. melanogaster).
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