RT-qPCR of LV tissue was performed to determine the transcript abundance of human AC8, genes that mediate neural autonomic input to LV (n=4 WT and 4 TGAC8 mice) and to detect genes regulating cytokines level in the heart (n=6 in WT and TGAC8). RNA was extracted from left ventricular myocytes (VM) with RNeasy Mini Kit (Qiagen, Valencia, CA) and DNAse on column digestion. The cDNA was prepared using MMLV reverse transcriptase (Promega). RT-qPCR was performed using a QuantStudio 6 Flex Real-Time PCR System (Thermo Fisher Scientific) with a 384-well platform. The reaction was performed with a FastStart Universal SYBR Green Master Kit with Rox (Roche) using the manufacturer’s recommended conditions; the sizes of amplicons were verified. Each well contained 0.5 μl of cDNA solution and 10 μl of reaction mixture. Each sample was quadruplicated and repeated twice using de novo synthesized cDNA sets. Preliminary reactions were performed to determine the efficiency of amplification. RT-qPCR analysis was performed using the ddCt method. Primers were selected with Primer Express 3.0 software (Applied Biosystems). Full list of primers used for amplification provided in Supplementary file 1p .
Faststart universal sybr green master kit with rox
Manufactured by Roche
The FastStart Universal SYBR Green Master Kit with Rox is a reagent kit designed for real-time quantitative PCR (qPCR) analysis. It contains a pre-mixed solution of SYBR Green I dye, FastStart Taq DNA Polymerase, reaction buffer, and ROX reference dye.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using faststart universal sybr green master kit with rox
1
Quantitative RT-PCR Analysis of Cardiac Gene Expression
2
Quantifying Dopamine Receptor Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RT-qPCR of the hippocampus and SA tissue was performed to determine the transcript abundance of dopamine receptors (n=4 WT and 4 TG AC8 mice). RNA was extracted from the hippocampus with TRIzol™ Reagent (Thermo Fisher Scientific, Waltham MA) and DNAse on column digestion.
The cDNA was prepared using MMLV reverse transcriptase (Promega, Madison, WI) using 500 ng of total RNA per reaction. RT-qPCR was performed using a QuantStudio 6 Flex Real-Time PCR System (Thermo Fisher Scientific, Waltham MA) with a 384-well platform. The reaction was performed with a FastStart Universal SYBR Green Master Kit with Rox (Roche, Indianapolis, IN) using the manufacturer's recommended conditions; the sizes of amplicons were verified. Each well contained 0.5 μl of cDNA solution and 10 μl of the reaction mixture. Each sample was quadruplicated. Preliminary reactions were performed to determine the efficiency of amplification.
RT-qPCR analysis was performed using the ddCt method. Hprt was used as a housekeeping gene and results represented as Mean of Relative Quantification (RQ) normalized to Hprt +/-SD (8).
Primers were selected with Primer Express 3.0 software (Applied Biosystems).
The cDNA was prepared using MMLV reverse transcriptase (Promega, Madison, WI) using 500 ng of total RNA per reaction. RT-qPCR was performed using a QuantStudio 6 Flex Real-Time PCR System (Thermo Fisher Scientific, Waltham MA) with a 384-well platform. The reaction was performed with a FastStart Universal SYBR Green Master Kit with Rox (Roche, Indianapolis, IN) using the manufacturer's recommended conditions; the sizes of amplicons were verified. Each well contained 0.5 μl of cDNA solution and 10 μl of the reaction mixture. Each sample was quadruplicated. Preliminary reactions were performed to determine the efficiency of amplification.
RT-qPCR analysis was performed using the ddCt method. Hprt was used as a housekeeping gene and results represented as Mean of Relative Quantification (RQ) normalized to Hprt +/-SD (8).
Primers were selected with Primer Express 3.0 software (Applied Biosystems).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!