FR AB - Science Reference

ABRF 2001 ABSTRACTS
T5
Technology resource grant opportunities.
M.T. Marron1, M.A. Tingle1, N. Pearson2, M.J. Saunders3; 1NCRR,
NIH, 6705 Rockledge Drive, Bethesda, MD 20892-7965, 2CSR, NIH,
3Natl. Sci. Fndn.
Federal funding is critical for equipping university-based Biomolecular
Resource Facilities with state-of-the-art technologies. In this session, representatives
from two Federal agencies, NSF and NIH, with day-to-day responsibility
for administration of instrumentation grants, will discuss the review
and funding of their various competitive grant programs.
The tutorial session will be split into two parts. The first half of the session
will provide an overview of the various instrumentation programs at NSF and
NIH of interest to ABRF members. The second half of the session will focus
on the preview processes at both agencies. Discussion will focus on practical
tips on how to prepare a successful NSF and NIH proposal. Formal presentations
will be followed by a question and answer period.
T7
Advances in DNA sequencing.
L.S. Hall1, E. Thomas1, K. Lilley2, G. Grills1; 1Albert Einstein Col. of Med.,
1695 Poplar Street, NewYork, NY 10464, 2Cambridge Univ.
This tutorial will review past advances in DNA sequencing and present new
techniques for sequencing difficult templates. Significant advances have
taken place in DNA sequencing in the past four years by virtue of improved
instrumentation, advances in the sequencing chemistries and alternative reaction
formats for sequencing difficult clones. In particular, the replacement of
Rhodamine terminators with Big Dye terminators increased the range of signal
strengths compatible with good data and eliminated many of the peak
height errors associated with rhodamine terminators; such as base calling
ambiguities linked to the pattern of a small ‘G’ following an ‘A’.
The recent introduction of dGTP terminators has improved the quality of data
from templates with high GC content or regions of secondary structure.
Data will be presented demonstrating the effectiveness of dGTP compared
to Big Dye. In addition, the efficacy of betaine (NNN Trimethylglycine) versus
DMSO will be discussed. New data will be presented demonstrating that
a combination of Big dye and dGTP can sequence through regions of high
G-C content without incurring compressions, a disadvantage of dGTP terminators
by themselves.
Finally, studies are in progress to evaluate the ability of different techniques
to sequence through various types of difficult motifs by addition of commercial
additives, such as the new GIBCO Rx Enhancer buffers. In general
difficult motifs, such as repeats, induce stalling of Taq and loss of signal
beyond the offending region. Depending upon the type of template, these
buffers can restore the processivity of the enzyme and resolve regions of secondary
structure. Results will be presented from studies that identify combinations
of additives that work well with particular motifs.
TUTORIAL ABSTRACTS
230 JOURNAL OF BIOMOLECULAR TECHNIQUES, VOLUME 11, ISSUE 4, DECEMBER 2000
T6
Advances in DNA sequencing.
M.A. Robertson; Univ. of Utah, 4A 438 School of Medicine,
50 N. Medical Drive, Salt Lake City, UT 84132
The Human Genome Project has been a major driving force for the development
of new instrumentation and methodology for high throughput
sequencing. During the last five years we have seen major advances both in
instrumentation and sequencing chemistries. This tutorial will present user
insight into one of the newer capillary instruments that promises to fit well
into a core laboratory setting. The tutorial will also look at the changes in sequencing
chemistry and present some solutions for difficult templates using
these new chemistries. This tutorial is designed to be interactive. There will
be a 30 minute question/answer session after the two main speakers, with a
panel comprising the speakers and members of the DNA Sequencing Research
Group. Please come armed with your most difficult sequencing problems
and help spark a lively discussion session.
T8
Incorporating a 16 capillary electrophoresis DNA sequencer
into a core facility.
P.T. Morrison; Dana-Farber Cancer Inst., 44 Binney St., JFB216,
Boston, MA 02115
In the past few years the Human Genome Project (HGP) has driven innovation
in DNA sequencing chemistry and instrumentation. For the most part this
has been beneficial for all types of DNA sequencing projects both large and
small. Recently manufacturers are now filling a void for instrumentation that
is better suited for facilities that sequence less than 100,000 templates per
year.
The incorporation of an Applied Biosystems Model 3100 Genetic Analyzer
into a core facility environment will be discussed.