Nebane N Miranda, Coric Tatjana, McKellip Sara, Woods LaKeisha, Sosa Melinda, Rasmussen Lynn, Bjornsti Mary-Ann, White E Lucile
High Throughput Screening Center, Southern Research, Birmingham, AL, USA
Department of Pharmacology and Toxicology, University of Alabama at Birmingham, Birmingham, AL, USA.
J Lab Autom. 2016 Feb;21(1):198-203. doi: 10.1177/2211068215620346. Epub 2015 Dec 9.
The development of acoustic droplet ejection (ADE) technology has resulted in many positive changes associated with the operations in a high-throughput screening (HTS) laboratory. Originally, this liquid transfer technology was used to simply transfer DMSO solutions of primarily compounds. With the introduction of Labcyte's Echo 555, which has aqueous dispense capability, the application of this technology has been expanded beyond its original use. This includes the transfer of many biological reagents solubilized in aqueous buffers, including siRNAs. The Echo 555 is ideal for siRNA dispensing because it is accurate at low volumes and a step-down dilution is not necessary. The potential for liquid carryover and cross-contamination is eliminated, as no tips are needed. Herein, we describe the siRNA screening platform at Southern Research's HTS Center using the ADE technology. With this technology, an siRNA library can be dispensed weeks or even months in advance of the assay itself. The protocol has been optimized to achieve assay parameters comparable to small-molecule screening parameters, and exceeding the norm reported for genomewide siRNA screens.
声学液滴喷射(ADE)技术的发展给高通量筛选(HTS)实验室的操作带来了许多积极变化。最初,这种液体转移技术仅用于转移主要化合物的二甲基亚砜溶液。随着具备水性分配能力的Labcyte Echo 555的推出,该技术的应用范围已超出其最初用途。这包括转移许多溶解于水性缓冲液中的生物试剂,如小干扰RNA(siRNA)。Echo 555非常适合用于siRNA分配,因为它在低体积时很精确,且无需逐步稀释。由于无需使用吸头,消除了液体残留和交叉污染的可能性。在此,我们描述了南方研究所高通量筛选中心使用ADE技术的siRNA筛选平台。借助该技术,siRNA文库可在实验本身进行前数周甚至数月进行分配。该方案已得到优化,以实现与小分子筛选参数相当的实验参数,且超过全基因组siRNA筛选所报道的标准。