Low cost, highly efficient library preparation
Echo Liquid Handlers enable library preparation in low microliter volumes for a range of sequencing methods including Sanger, 16S, and Next-Generation Sequencing (NGS) – to dramatically reduce reagent costs, save samples, and eliminate steps — all while improving library quality and throughput.
With Sanger sequencing, BigDye reagent costs can be a limiting factor. Sequencing volumes can be reduced from 10 µL to 2.5 µL with the use of the Echo Liquid Handler for a 4-fold reduction in cost per reaction.
16S rRNA sequencing has long been the method of choice for microbiome researchers. The use of the Echo Liquid Handler allows for miniaturized 16S rRNA sequencing, and NGS reagent cost reduction also enables researchers to shift to miniaturized shotgun sequencing.
Use of the Echo Liquid Handler for NGS library preparation versus conventional liquid handlers allows for up to a 100-fold reduction in reaction volumes, dramatically reducing study reagent costs.
|Manual Pipetting||Echo® Liquid Handler|
|Amount of DNA||50 ng||0.06 – 2.0 ng|
|DNA volume (Rxn)||25 µL||200 nL|
|Library prep volume (Rxn)||25 µL||300 nL|
|Total volume||50 µL||0.5 µL|
|Reactions per kit||96||9600|
|Cost per reaction||$72.91||$0.73|
When NGS sequencing programs scale, the cost of library preparation is a limiting factor. With the Echo Liquid Handler's low-volume acoustic transfer capability, researchers can significantly increase the number of libraries generated while maintaining current budgets.
“By using the Echo 525, a scale-down by 50 – 100x can be achieved with minor modifications of the protocol, leading to significant cost reduction without sacrificing data quality. Furthermore, we demonstrate the superiority of contact-free liquid transfers in molecular biology applications with regard to reproducibility and contamination prevention.”
Stephan Lorenz | Wellcome Trust Sanger Institute, UK
Echo Liquid Handlers can rapidly pool and normalize oligos or DNA libraries from any well of a microplate. With average transfer times of no more than a few seconds per well, Echo Liquid Handlers can reduce pooling times by more than 80% compared to traditional liquid handlers. Since Echo systems can transfer volumes as little as 25 nanoliters, high concentration libraries do not have to be diluted prior to transfer. This capability results in simultaneous normalization while pooling. Furthermore, by avoiding tip-based sample dilution, the propagation of errors from sample retention by tips is eliminated. This drastically improves the concentration accuracy of all libraries pooled.
FEATURED TECH NOTE
Jefferson Lai, et al.
As the cost of sequencing has continued to decline by orders of magnitude in the past ten years, scientists are enabled to ask deeper and more complex questions about transcriptomics. Consequently, the variety, quantity, and demands of RNA sequencing experiments have all increased. The Clontech SMART-Seq® v4 Ultra® Low Input RNA Kit incorporates the SMART (Switching Mechanism at 5’ End of RNA Template) technology. This technology utilizes the template switching activity of reverse transcriptases to add PCR adapters directly to both the 5’ and 3’ ends of the first-strand cDNA before amplification using LD PCR. Here, we document information and results pertaining to the miniaturization of this process utilizing the Echo® 525 Liquid Handler, effectively reducing the reagent cost and processing time for the workflow, addressing current throughput needs of RNA sequencing.