A new method for generating arrayed RNAi screening tools for any organism
Poster May 08, 2018
Jesse Stombaugh, Megan Basila, Amanda Haas, Annaleen Vermeulen, Melissa L. Kelley, Anja van Brabant Smith
For studies in alternate species, siRNAs needed to be custom synthesized at a relatively high cost and very long manufacturing time. To improve the accessibility of these large screening libraries to researchers working in non-human or mouse model systems, we have recently developed a new cost-effective enzymatic manufacturing method to generate large collections of siRNAs, called Dharmacon™ Zoonome™ siRNAs (z-siRNAs). Like other well-established siRNA product lines, they are designed using the proprietary SMARTselection design algorithm for potent and specific gene expression knockdown, but are manufactured on-demand using a novel enzymatic method. Now available are customized arrayed collections of pools of four z-siRNAs to each gene target for screening hundreds or thousands of genes for any organism with an annotated genome. Here we show successful gene knockdown in Cricetulus griseus (Chinese hamster) and Bos taurus (bovine) cell lines using Zoonome siRNAs. With this new availability of large screening libraries, the power of unbiased functional screening in nearly any species can accelerate findings important to animal health, food safety, and infectious disease.
When there is a need to quickly analyze samples using a number of different PCR assays, it is likely that optimal conditions for each assay will not be the same. First, different assays often will require different annealing temperatures for their primers. In addition, amplicons may be designed to be of different lengths and therefore require varying durations of the extension step.READ MORE