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Research Project
My project involves the development of microarrays. I am currently
working with Dr. LaPres who obtained the NIA 15K clone set.
The clones are first PCR-amplified. They are then precipitated
and transferred onto 384-well plates for microarray slide printing.
The objective of Dr. Zacharewski's lab is to identify the genes
that are expressed in mouse testis and develop mouse testis-specific
microarrays. As the NIA 15K microarrays become available, they
will be screened for the genes expressed in mouse testis through
hybridization experiments and data analyses.
My other job is to prepare 12 genes to be put on the mouse
testis-specific microarrays whose IMAGE clones are not available.
Primers had been designed to amplify a 200~300bp part of each
gene and ordered prior to my arrival. RT-PCR is done on RNA
extracted from various mouse tissues. The RT-PCR products are
then digested and cleaned for subcloning where they are ligated
and transformed into the pGEM vector and XL1 blue cells, respectively.
The cells are then plated, inoculated, and DNA-prepared to check
for the insert. The presence of correct insert is then further
verified by sequencing the insert and BLASTing the sequence
to that in the NCBI database.
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