Amplification
It's nice to be working from home. I have the radio on, a pot of tea, a Lindt bunny, and a cat. All are distracting from the qPCR results in front of me.
I haven't done PCR in over 2 years. I did my first run on the Applied Biosystems ABI Prism 7000 machine in Pharmacology today, looking for expression of cannabinoid receptor 1 and oxytocin receptor mRNA in rat hypothalamus and dorsal striatum respectively. The setup was quicker due to using commercial master mixes (yay!). The software interface is a little daggy but simple enough. But my endogenous control, 18S ribosomal subunit, appears to be varying in its expression, which it shouldn't be. I need to troubleshoot this.
There are numerous gene quantification advice websites full of quite reliable information. But I need to re-hone my understanding of the kinetics of the PCR reaction and the influence of various parameters. Since chaos reigns and the sensitivity of many systems to slight variations in initial conditions is crucial, the many places to compound error in the process of quantitative PCR have to be accounted for, from extracting tissue from the animal to final analysis.
I haven't done PCR in over 2 years. I did my first run on the Applied Biosystems ABI Prism 7000 machine in Pharmacology today, looking for expression of cannabinoid receptor 1 and oxytocin receptor mRNA in rat hypothalamus and dorsal striatum respectively. The setup was quicker due to using commercial master mixes (yay!). The software interface is a little daggy but simple enough. But my endogenous control, 18S ribosomal subunit, appears to be varying in its expression, which it shouldn't be. I need to troubleshoot this.
There are numerous gene quantification advice websites full of quite reliable information. But I need to re-hone my understanding of the kinetics of the PCR reaction and the influence of various parameters. Since chaos reigns and the sensitivity of many systems to slight variations in initial conditions is crucial, the many places to compound error in the process of quantitative PCR have to be accounted for, from extracting tissue from the animal to final analysis.