Trevor L. Hawkins, Cheryl Evans, David Goon, Jarrod Loncor, Tara O'Connor, Jim McDermott, Niall Moloney, Meghan Lane, William Fitzhugh, David Wang, and William Lee.
Whitehead Institute/ MIT, Center for Genome Research, One Kendall Square, Cambridge, MA 02139.
There is a major need for the development of a system which can accomplish the integrated tasks of DNA isolation and proceed with purification and the setup of sequencing reactions. Here we demonstrate the feasibility of such a system from both a biochemical and engineering perspective.
We have set up collaborations with CRS robotics corp, Packard Instruments, Tecan US and Techne Inc. to design and construct a 'factory style' laboratory system which is called the Sequatron.
The major component of our system is an articulated CRS 255A robotic arm which is track mounted. The deck of the robot contains several new/modified XYZ robotic workstations, a novel thermal cycler with automated headed lids, carousels and custom built plate feeders.
Biochemically, we have employed our Solid-phase reversible immobilization (SPRI) technique to isolate and manipulate the DNA throughput the process. Specifically we have set up the Sequatron to isolate DNA from M13 phage or crude PCR products using the same protocol and procedures. From M13 phage we obtain approximately 1ūg of DNA per well which is sufficient for multiple sequencing reactions.
The current throughput of the system is 80 microtiter plates of samples from M13 phage supernatants or crude PCR products to sequence ready samples every 24 hours. Recently, new enzymes, new energy transfer primers and higher density microtiter plates have opened up possible increases to in excess of 25,000 samples per 24 hour period.
Supported by a grant from the Office of Health and Environmental Research of the U.S. Department of Energy under contract DE-FG02-95ER62099.