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Research Project
As a Dalhousie University undergraduate co-op student, I have
been involved primarily in two projects in the Zacharewski lab.
First, I have been assigned to work under the supervision of
Kirsten Fertuck, whose project involves the use of cDNA microarrays
to monitor the expression of thousands of genes in estrogen-responsive
tissues. It has been my responsibility over the summer to prepare
the last 250 clones of estrogen-responsive genes for printing
onto a custom microarray. This first involves the sequence verification
of the clones, obtained from the IMAGE consortium. Sequencing
is expensive and it would be wasteful to sequence clones that
can be identified as being contaminated and thus discounted
by inexpensive and routine methods. The plasmids are prepped
and digested then are qualitatively analyzed by agarose gel
electrophoresis. Clones that seem to contain only one plasmid
are sequenced, while clones that seem to contain more than one
plasmid are streaked on agar plates for clonal isolation to
ensure that pure DNA is sent to the sequencing facility. When
the samples are sequenced, the BLAST tool on the NCBI website
is used to compare the accession number for the ordered clone
with the in-house derived sequence to determine if it was the
correct clone. If so, the clone is PCR amplified and spotted
on the array plate.
My laboratory experience also includes work in Dr. John LaPres’
lab, where I am involved in the preparation of clones from the
NIA 15K clone set for printing onto microarray slides. These
clones have already been sequence verified and therefore need
only to be PCR amplified and spotted onto the array plates.
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