In this final installment on how to improve your qPCR gene expression profiling (see 6 Tips to Improve qPCR: part 1 and part 2 for previous discussion), we will discuss pipetting tips and the benefits of a separate reverse transcription step in qPCR template preparation.
Tip #5. Good Laboratory Practice
In this next installment on how to improve your qPCR gene profiling (see 6 Tips to Improve qPCR for previous discussion), we would like to discuss the limitations of melting curve analysis and PCR inhibitors.
Tip #3. Melting curve analysis is NOT sufficient for assessing qPCR product specificity.
In
Gene expression analysis is critical for understanding the transcriptome profiles of primary cells and how they directly influence the cells’ functionality. In addition, the transcriptome profile can affect cell proliferation, differentiation, senescence, or apoptosis. Traditional reporter-gene assays
Dye-based qPCR assays, such as SYBR® Green and EvaGreen®, have many advantages over probe-based qPCR assays. Dye-based systems are desirable because they are highly adaptable, cost effective, and time efficient. qPCR specificity, however, is a vital factor to consider when using dye-based qPCR assays.
