Monthly Archives: August 2018
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- 八月 29, 2018
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 qPCR validation. However, as essential as qPCR is, qPCR loading can be challenging.
Why is qPCR loading difficult? First, qPCR is typically performed using 96- or 384-well plates. The replicon template, primers and master mix (consisting of polymerase, buffer and dNTPs) must be properly loaded to individual wells. The sample loading step can be challenging because of the large number of wells and it is extremely easy to introduce errors when loading. Second, when quantifying low copy number genes, white qPCR plates are preferred over transparent ones, as several studies have shown that white plates can offer better qPCR sensitivity. The opacity
- 八月 20, 2018Cell-based assays are widely used in basic and translational research as cost-effective and accessible models to mimic in vivo responses. To obtain reliable data, assessing the health of cultured cells prior to any assays is highly recommended. Furthermore, many cell-based assays require quantification of cell growth. Cell health and growth can be determined by quantifying cell viability, proliferation, or apoptosis.
Below, we compare some commonly used assays to help you determine which type is suitable for your experimental design.
- Cell viability assays enumerate the ratio of live and dead cells in a population. Cell viability can simply be achieved by staining and counting live or dead cells. A deeper assessment of cell health can be attained by measuring cell metabolic activity, such as the ability to reduce tetrazolium salts in MTT and WST-1 assays.
- Cell proliferation assays assess dividing cells. Some common assays include BrdU incorporation (BrdU Assays)
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