Monthly Archives: September 2017
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- 九月 21, 2017
Next generation sequencing (NGS), such as Whole Exome Sequencing (WES) and RNA-Sequencing (RNA-Seq), provides a high throughput approach for DNA sequencing and gene expression analysis. It is rapidly advancing our knowledge on almost all aspects of genetic research and shedding light on how individual or groups of genes may regulate biological processes. Powerful as NGS is, however, many high impact journals require that NGS data include qPCR validation.
So why is qPCR validation still such an important step? There are 3 main reasons.
First, compared to qPCR, NGS is considerably more complicated and consequently the potential for errors is increased. Reproducibility can also be a problem due to the complexity of NGS experiments.
Second, there can be intrinsic biases in NGS. For one, due to the random sampling nature of NGS, the sensitivity is largely based on the "sequencing depth", or how many times the template gets read. If the transcripts are expressed at low levels, it may not reach
