( Lynn Jordan James Batchelor...)Automation of the Eppendorf Perfectprep BAC 96 DNA Kit on the Caliper Life Sciences Sciclone ALH 3000

Lynn Jordan, James Batchelor, and Ed Alderman
Applied Science and Technology Department, Caliper Life Sciences Inc., Hopkinton, MA 01748

Kelly M. Clark, Joseph A. Hensley and Robert P. Weisbein
Brinkmann An Eppendorf Company, Westbury, NY

Jennifer L. Halcome and George R. Halley
Eppendorf 5 Prime, Inc. Boulder, CO

Introduction

To assemble large genome constructs, Bacterial Artificial Chromosomes (BACs) are necessary for fingerprinting mapping and BAC-end sequencing. It is critical to have an automated method to obtain high quality BAC DNA. We have completed feasibility studies integrating Eppendorfs Perfectprep BAC 96 reagent kit with Caliper Life Sciences Sciclone ALH 3000 to achieve high quality BAC in a timely fashion. A 96- channel pipetting head performs reagent additions using disposable tips. An on-deck shaker is used to resuspend the pelleted material, and is used for mixing in other steps. Positive pressure is used to perform all filtrations steps. Through the use of an on-deck gripper, plates can be relocated on the deck to provide a Hands-Off automated solution.

Introduction

Purification of sufficient quantities of high quality BAC DNA, when using lower volumes of reagent and starting culture, has posed a challenge for researchers. Eppendorf has solved this problem with the development of the Perfectprep BAC 96 that is capable of processing 96 samples in a microplate format. Figure 1 shows an overview of the Prefectprep BAC 96 process. For many researchers, projects involve the processing of thousands of samples to acquire the downstream data required. In an effort to achieve low- to mid- throughput purification (1-16 plates in an eight-hour workday), we have performed initial experiments to integrate the Eppendorf Perfectprep BAC 96 kit onto the Caliper Life Sciences Sci
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