Effect of Liquid Handling Quality Control on Biological Assay Performance
Poster Feb 27, 2012
Nathaniel Hentz, Ph.D and Tanya Knaide
Molecules from a high throughput screening (HTS) campaign are often selected for further development after a single interrogation. HTS groups are also constantly challenged with improving efficiency by incorporating new technologies and complex biology. Hence, in order to obtain promising candidates, high quality assays are required. Assay variability is an important quality attribute and is a combination of liquid handling, biological and random variability. Liquid handling variability is often underappreciated, yet can have a huge impact on the outcome of an assay. Many laboratories rely on only precision to estimate the quality of the liquid handling. Optimization of liquid handling is so critical to improve the assay’s performance that precision and accuracy must both be assessed. This study was designed to examine (1) liquid handling quality control (QC) tools and (2) the effect of liquid handling variability on a model biological assay.
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE