qPCR is a powerful tool for quantification of gene expression levels and copy number variation. Despite the advances in next-generation sequencing (NGS), qPCR still serves as the "gold standard" for gene expression analysis. Due to poor reproducibility and vast lab-to-lab variation, all NGS data requires
The 2017 Nobel Prize in Physiology or Medicine was awarded to Jeffrey C Hall, Michael Rosbash, and Michael W Young for research that established key mechanistic principles on how circadian rhythms are regulated. Circadian rhythms are endogenous oscillations adjusted to changing external cues and driven
Protein-coding DNA, or “functional” genes, only accounts for up to 2% in the human genome. Until recently, the remaining 98% have been referred to as “junk DNA” for their putative noncoding feature. The name itself implies that junk
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
Despite being essential for any experiment involving DNA replication, people rarely give primers a second thought. To coincide with the launch of our new qPCR gene reference tool, we’ll be giving them a little more of the praise they deserve.
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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.
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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
qPCR array provides a quick, powerful and sensitive approach for gene expression profiling. To help our customers acquire accurate and consistent results, we are happy to share some tips for better qPCR.
Tip #1. Good primer design is the key to a successful qPCR array.
The success of a qPCR array depends
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.
