Purification Biomek® 2000
Beckman
Coulter Application (system and method support)
Automation of Total RNA from Cell Cultures on the Biomek® 2000 Automation Workstation using Promega's SV 96 Total RNA Isolation System
Matthew Cu**, Dana P. Campbell**, Hobert Wai** and Keith Roby**
Beckman Coulter, Inc.
**Indianapolis, IN and ***Fullerton, CA
Terri Grunst and Daniel Kephart
Promega Corporation –
www.promega.com
To satisfy the need for the isolation of human total RNA from tissue
and cell culture we have developed a rapid and fully automated method
for the isolation of total RNA using the Promega SV 96 Total RNA Isolation
System (Promega, Inc., Madison, WI) and the Biomek® 2000 Laboratory
Automation Workstation from Beckman Coulter. Total RNA is purified from
cell lysates using a 96-well vacuum filtration procedure, eliminating
the need for centrifugation. Washing the bound RNA does not require disassembly
of the vacuum manifold; filtrate waste products are delivered directly
to a vacuum trap. A direct DNase treatment step on the SV 96 Binding plate
substantially reduces genomic DNA contamination. Total RNA is collected
in a convenient, 96-well plate, making the process simple and easy to
implement in the RNA research laboratory. Furthermore, the method is capable
of complete automation of the isolation and purification of 96 high-quality
total RNA samples in less than 50 minutes.
The isolated total RNA was evaluated for quality, reproducibility, degradation,
and cross-contamination using gel electrophoresis and RT-PCR.
* All trademarks are the property of their respective owners. Where applicable, the PCR process is covered by patents owned by Roche Molecular Systems, Inc., and F. Hoffman-LaRoche, Ltd.
