Total RNA from liver and inoculated tumors was isolated using Trizol® Reagent (Life Technologies, Grand Island, NY) and purified using Qiagen RNeasy kit (Valencia, CA) following manufacturer’s protocols. Quantity and quality of the RNA was assessed with the Nanodrop® ND-1000 UV-Vis Spectrophotometer (Nanodrop Technologies, Wilmington, DE). RT-qPCR analysis was performed with total RNA using the ABI Prism 7700 and TaqMan EZ RT-PCR core reagent kit (Applied Biosystems, Grand Island, NY) with pre-made mouse-specific TaqMan gene Expression Assays for Bmal1 (Mm00500226_m1), Per2 (Mm00478113_m1) and 18S rRNA (Mm03928990_g1), which was used as the reference gene. Relative gene expression was calculated using ΔΔCt method as previously described (Panda et al., 2002 (link)).
Taqman ez rt pcr core reagent kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The TaqMan EZ RT-PCR core reagent kit is a tool used for reverse transcription and real-time PCR (RT-PCR) amplification of RNA samples. It contains the necessary reagents and enzymes to perform both the reverse transcription and the subsequent PCR reaction in a single tube.
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Lab products found in correlation
Abi prism 7700, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Nanodrop nd 1000 uv vis spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Rneasy kit, by Qiagen (1 mentions)
Abi prism 7900 sequence detector system, by Thermo Fisher Scientific (1 mentions)
2 protocols using taqman ez rt pcr core reagent kit
1
Liver and Tumor RNA Isolation and RT-qPCR Analysis
2
Real-time RT-PCR for Norovirus Genotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The extracted nucleic acids were screened to identify the genogroup (GI and GII) by real-time RT PCR reactions as described previously [15 (link)]. The reactions were set up using the TaqMan® EZ RT PCR Core Reagent kit (Applied Biosystems, Foster City, CA, USA). All reactions were carried out in ABI PRISM 7900 Sequence Detector System and the results were analyzed with Sequence Detection Software version 2.1 (Applied Biosystems, Foster City, CA, USA).
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