FR AB - Science Reference

Implementation of Automation
in a Small-Scale DNA
Sequencing Core Facility
Jennifer L. Ware, Laurie Moran,
Ching-Lun Lin, and Barton Slatko
New England Biolabs, Inc., Beverly, MA
METHODS
REVIEWS
New England Biolabs (NEB) sequencing core facility provides
automated sequencing services to support various
company-wide projects in house, but on a very small scale of
about 1000 to1500 reactions per month. A procedure has
been implemented at the NEB core sequencing facility to
integrate simplified methods and robotics to provide a more
efficient small-scale process. This has been done using a
Beckman Biomek 2000 robot combined with an MJ DNA
Engine, 96-well plate cycler (PTC-200), AB 373 and 377
sequencers, BMA Singel gels, and several other materials that
help reduce the time required for otherwise lengthy procedures
in a cost-efficient manner. Protocols have also been
developed for efficient sequencing of a variety of templates
submitted to the NEB core facility. (J Biomol Tech 2000;11:
151–154)
KEY WORDS: automation, robotics, sequencing template
cleanup, small-scale DNA sequencing.
ADDRESS CORRESPONDENCE AND REPRINT REQUESTS TO: Jennifer L.
Ware, New England Biolabs, Inc., 32 Tozer Road, Beverly, MA
01915 (email: ware@neb.com).
&
Journal of Biomolecular Techniques
11:151–154 © 2000 ABRF
RF AB
Genome sequencing and analysis technology
has advanced in a manner that allows companies
to generate enormous amounts of data
per day, via automated processes, in an effort to complete
genome projects and use that information as a
source of possible disease treatment and control.
Unfortunately, smaller core facilities, which have less
of a need to produce such high-throughput data, are
not the primary targets of application of these
advanced technologies and are in need of simplified
methods. The core sequencing laboratory at New
England Biolabs, Inc. (NEB, Beverly, MA) has combined
recent advances in sequencing technology to
efficiently simplify the process from beginning to end.
The sequencing core facility provides additional measures
of quality control of appropriate DNA products
for sale at NEB, such as sequencing primers and other
oligonucleotides, cloning vectors by sequencing reaction
assay, and vector and phage display products to
ensure correct sequence. For our in-house research
program, sequences are required from vector and
overexpression constructs of clones, cDNA, polymerase
chain reaction (PCR), phage display, and other
sequencing templates, including our involvement in
the Filarial Genome Project (http://www.neb.com/
fgn/filgen1.html). Among those templates submitted
for analysis are plasmids, M13 derivatives, � phage,
PCR products, cosmids, and bacterial artificial chromosomes
(BACs), each of which requires a different
amount of template and/or primer per reaction and
sometimes different sequencing reaction conditions as
well.
ELECTRONIC SUBMISSION
To make this process efficient from beginning to end,
researchers start by submitting an electronic form to
the core facility in which key information necessary to
perform the sequencing is provided. The form is accessible
on the DNA Sequencing Core Facility intranet
JOURNAL OF BIOMOLECULAR TECHNIQUES, VOLUME 11, ISSUE 4, DECEMBER 2000 151