GATCAAAACCATCAGAATCCCTTTGGTTGATGAATTGGAAACGTACAAAGAAATCACGAGAATCATCAACCTCTTGCGCTCAGAAAACATTTCGATT
CAAGCTGCAACCTCGATTGCGTACTAGGAATTGCATATTATGGGAGAAAAGAAGCGATTGAGAACCAGCAGCAACCTAGCAATTACTTTCCGTTAA
GCACCCATCCCAACCCTTTCTTTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCATTTCCTATCTCCTTACTGTCACATCCTCACCAAATCCATCTTCC
TCCTCACCTCCATCCTCATCCTAATCATTTGCAGCCTCTTTCCTCTTAACACCTCGATTCCTTCGAACCCGTTACTGTCACCTCTATCCAACAAGATC
DNA SEQUENCING
SERVICES
GENETIC LABORATORY
Source: US Department of Energy Human Genome Project
For further information on the DNA
sequencing services, please contact:
MS ISO 9001:2000 CERTIFIED
Dr. Norwati Muhammad
Tel.: (+603) 62797144
Fax: (+603) 62797856
E-mail: [email protected]
Dr. Lee Soon Leong
Tel.: (+603) 62797145
Fax: (+603) 62797856
E-mail: [email protected]
Or visit our website at:
http://www.frim.gov.my
FOREST RESEARCH INSTITUTE MALAYSIA
52109 KEPONG, SELANGOR DARUL EHSAN
MALAYSIA
DNA SEQUENCING
SERVICES
OVERVIEW
The
DNA Sequencing Services at the Genetic
Laboratory of Forest Research Institute Malaysia
(FRIM) is currently available using Applied
Biosystems 36-lanes Model 377 Automated DNA
Sequencer. The ABI Sequencer system is an
electrophoresis detection system that automates
the separation of fragments generated during
sequencing reactions and subsequent base-call
determinations.
A
Sanger-based
dideoxy
sequencing strategy involving the incorporation
of fluorescent dye-labeled terminators into the
sequencing
reaction
products
is
used.
Fluorescent dye labels (a different colour for
each of the four nucleotide bases) allow
sequences to be determined within a single lane
on the gel. During data collection, the sequence
reaction products are detected as it
electrophoreses passes scanning argon laser
positioned near the bottom of the gel. The laser
beam excites the fluorescent dyes causing them
to fluoresce. This emitted fluorescence is
collected and passed sequentially through four
filters and then directed onto a photo multiplier
tube. The digitised output is automatically
tracked but the result is always checked
manually. At the end of the data preprocessing,
an analysis software package converts the raw
data into sequence information.
In our laboratory, we use Big Dye
Terminator chemistries. This chemistry is able to
increase the sequencing of a wide variety of
templates successfully, including those that
contain regions of secondary structure.
We have procedures to analyse both single
and double strand DNA samples. Currently, we
provide the M13 forward and reverse, SP6, T3,
T7, and SK promoter primers for sequencing
reactions at no additional charge. The average
read length for this service is between 500 - 700
base pairs, depending on template quality.
Our goal is to help you with DNA sequencing
by providing reasonably priced and high quality
service.
WHAT ARE REQUIRED
•
•
Good purity and integrity of the template
DNA
If you are supplying your own primers, they
should have a Tm between 50°C and 65°C,
and lack potential secondary structure.
POLICIES
1. All samples are processed on a first come
first served basis.
2. Concentration of all DNA templates will be
quantified using spectrophotometer and/or
electrophoresis to ensure its adequacy.
Unsuitable samples will not be processed
(customers will be notified for resubmission
of new DNA templates).
3. Each set of sequencing reactions includes at
least one positive control reaction.
4. Sequencing that fail to meet quality
standards will be repeated.
5. All sequencing
confidential.
results
will
be
kept
CHARGES
•
RM50.00/template
SERVICE HOURS
DNA samples for analysis should be delivered to
the FRIM Genetic Laboratory during the
following hours:
• Monday - Friday
9.00 a.m. - 12.00 p.m.
2.30 p.m. - 4.00 p.m.
Customers can expect to receive their results in
approximately 2 weeks.
WHAT WE WILL PROVIDE
Customers will receive the results as a sequence
file and a colour electropherogram for each
template. Data can be provided in print-out
form, on floppy disk, CD, or can be sent by email upon request.
DATA ANALYSIS AND EDITING
Free software for viewing and editing DNA
sequencing electropherograms is available. To
view the DNA sequence electropherograms you
can use Chromas for Microsoft Windows. The
Chromas homepage has more information on
this software. An older version, 1.45, is available
as freeware.
Feb, 2006