Total RNA was isolated using Trizol/RNEasy reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. Isolated RNA was precipitated in isopropanol, washed in ethanol, and dissolved in RNase-free water. RNA concentration was determined by measuring absorbance at 260 and 280 nm. Total RNA was reverse transcribed using the TaqMan Gold RT-PCR kit (Applied Biosystems, Foster City, CA). Amplification was performed either utilizing the Universal TaqMan master mix and primer/probe sets specific for the genes of interest (Applied Biosystems) and examined on a Lightcycler 480 II (Roche), or alternatively amplified using the SYBR Green Master Mix (Applied Biosystems, N8080234 and 4309155) and examined on a 7500 Real-Time PCR system (Applied Biosystems), as previously described [28 (link)]. A purchased human B cell (CD19+) total RNA was used to establish a baseline for comparison between mRNA levels in CLL and a healthy subset (Miltenyi). Quantitative, real-time RT-PCR was performed by the relative quantification (2−ΔΔCt) method using β-actin as the reference mRNA.
Universal taqman master mix and primer probe sets specific for the genes
Manufactured by Thermo Fisher Scientific
The Universal TaqMan master mix is a ready-to-use solution designed for real-time PCR applications. It contains all the necessary components, including DNA polymerase, dNTPs, and buffer, to perform TaqMan-based gene expression analysis. The primer/probe sets are specific for the genes of interest and are designed to work with the Universal TaqMan master mix. These products provide a streamlined and standardized approach for reliable and consistent gene expression studies.
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Lab products found in correlation
Taqman gold rt pcr kit, by Thermo Fisher Scientific (2 mentions)
Trizol rneasy reagent, by Thermo Fisher Scientific (2 mentions)
7500 real time pcr system, by Thermo Fisher Scientific (2 mentions)
Sybr green master mix, by Thermo Fisher Scientific (2 mentions)
Lightcycler 480 2, by Roche (2 mentions)
2 protocols using universal taqman master mix and primer probe sets specific for the genes
1
Quantitative RT-PCR Analysis of CLL
2
Quantitative RT-PCR Analysis of CLL
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Total RNA was isolated using Trizol/RNEasy reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. Isolated RNA was precipitated in isopropanol, washed in ethanol, and dissolved in RNase-free water. RNA concentration was determined by measuring absorbance at 260 and 280 nm. Total RNA was reverse transcribed using the TaqMan Gold RT-PCR kit (Applied Biosystems, Foster City, CA). Amplification was performed either utilizing the Universal TaqMan master mix and primer/probe sets specific for the genes of interest (Applied Biosystems) and examined on a Lightcycler 480 II (Roche), or alternatively amplified using the SYBR Green Master Mix (Applied Biosystems, N8080234 and 4309155) and examined on a 7500 Real-Time PCR system (Applied Biosystems), as previously described [28 (link)]. A purchased human B cell (CD19+) total RNA was used to establish a baseline for comparison between mRNA levels in CLL and a healthy subset (Miltenyi). Quantitative, real-time RT-PCR was performed by the relative quantification (2−ΔΔCt) method using β-actin as the reference mRNA.
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