omation mbers - Society for Laboratory Automation and Screening

TP033
Xingwang Fang
Ambion, Inc.
Research and Development
2130 Woodward Street
Austin, Texas 78744
xfang@ambion.com
High Throughput RNA Isolation – Methods Comparison
162
Co-Author(s)
Roy C. Willis
Quoc Hoang
Michael Siano
The demand for high throughput RNA isolation has been dramatically increasing with wide application of RNAi,
expression profiling and molecular diagnosis. We will present a comparison of various RNA isolation methods that
have been adapted to high throughput platforms, focusing on consistently high yield and quality of isolated RNA,
reduction of cross-contamination, and simplicity and robustness of the protocol. The most widely used method
for high throughput RNA isolation uses glass fiber filter plate where RNA binds to filter in the presence of high salt
and alcohol. New methods have been developed using microspheric beads. In general, microspheric bead-based
approach results in more consistent RNA recovery than glass fiber filter based RNA method. This is because
beads can be fully resuspended in solution to enable more thorough mixing, washing, and elution. In addition,
bead-based method is easier to automate than filter plate-based method where vacuum may cause crosscontamination
and occasional filter clog may fail all samples in the whole plate. We will show comparison results
on RNA isolation from cultured cells, as well as for viral RNA isolation from liquid biological samples. We will also
discuss a technology for generating cDNAs and performing PCR directly from cells. In this technology, nucleases
and reverse transcriptase inhibitors are deactivated during a simple lysis step. The resulting cell lysate can be used
directly to make cDNA and for one-step qRT-PCR, bypassing the RNA isolation step. Data from the application of
this technology to siRNA-mediated gene silencing will be presented.
TP034
W. Steven Fillers
TekCel, Inc.
103 South Street
Hopkinton, Massachusetts 01748
steve.fillers@tekcel.com
ENCOMPASS: A New Expandable Approach for Large-Scale, Multi-Format Sample
Management
Traditional approaches to automating large-scale sample storage, retrieval, and distribution have been limited
to costly, customer engineered “cathedral” systems. Though these systems have met the capacity and process
control demands of the select few, they present a number of significant implementation burdens and operational
risks to many. Large-scale systems (>1M samples) are typically engineered to meet fixed capacity and throughput
requirements, necessitating immediate capital investment to meet potential future storage needs. Custom
development and construction of facilities to house the system is frequently necessary, further expanding the
cost and time for system implementation that, by design, provides no cost-effective option for progressive
expansion. Additionally, due to the complexity of these single-point-of-access systems, failures of individual
robotic components can bring the entire sample management operation to a halt. To address these challenges
TekCel has developed Encompass, an expandable, large-scale sample storage and retrieval system configurable to
specific customer requirements. Encompass employs a versatile system architecture based on TekCel’s Universal
Storage Module (USM). USMs serve as functional building blocks combined to provide the capacity and format
scope (plates, tubes, vials, racks) required by the customer’s operational needs. Standard environmental control
and robotic elements, established in TekCel’s modular sample management systems (TubeStore, PlateStore, etc.),
are utilized to meet desired throughput and sample processing specifications while maximizing sample integrity.
Encompass systems provide the highest degree of flexibility to address customer workflow requirements and allow
staged implementation to support process change and manage capital outlay.