omation mbers - Society for Laboratory Automation and Screening

WP021
Dan Kephart
Promega Corporation
Scientific Applications
2800 Woods Hollow Road
Madison, Wisconsin 53711
dkephart@promega.com
Automated Genomic DNA Purification From Large Volumes of Whole Blood
202
Co-Author(s)
Cris Cowan
Terri Grunst
We have developed a novel reagent/hardware system that enables completely automated and scaleable genomic
DNA isolation from up to 10 ml of whole blood.The method takes advantage of the unique properties of Promega’s
MagneSil paramagnetic particles in conjunction with a novel tube rack/magnetic device to allow isolation using
standard 50ml conical tubes on a variety of automated platforms. Greater than 300µg of DNA is isolated from
whole human blood containing a minimum of 1 x 10 6 white cells; over 500µg can be obtained from blood with
higher white cell counts. The purification procedure is completely automated and does not require differential lysis,
fractionation of white cells, centrifugation, precipitation, or rehydration of DNA prior to analysis. Performance of the
isolated DNA is demonstrated in real-time and endpoint PCR, restriction digest, and STR analysis.
WP022
Alan Kishbaugh
Meso Scale Discovery
9238 Gaither Road
Gaithersburg, Maryland 20877
akishbaugh@meso-scale.com
Co-Author(s)
Gisbert Spieles, Erin Grossi, Kent Johnson,
Rob Calamunci, George Sigal, Svetlana Leytner,
Jacob N. Wohlstadter
Multiplex Measurements of Cytokines in High Density Formats Using Multi-Array
Technology
This poster presents multiplexed immunoassays for human cytokines using Meso-Scale Discovery’s (MSD’s) Multi-
Array platform. These assays were conducted in MSD’s new Multi-Spot plates, using a sample kit provided by
MSD. The kit includes plates pre-coated with capture antibodies for each cytokine, a cocktail of labeled detection
antibodies and other related assay reagents. MSD’s multiplex cytokine assays have no-wash formats, rapid
protocols (2.5 hours) and fast read times (~70 sec. per plate). They are compatible with complex assay matrices
(e.g., cell supernatants, cell lysates, serum, blood). No-wash assays require as little as 10 µl of sample, and
have excellent selectivity (99.5%) and sensitivity (~1-10 pg/ml). The wide dynamic range (>3 logs) allows users
to avoid multiple dilutions. We present data from a panel of numerous cytokines. Specifically, we demonstrate
the performance of a 384-well Multi-Spot kit for measuring IL-1ß, IL-2, IL-6 and TNF-a. We also compare the
performance of this kit with other MSD Multi-Spot plate formats.