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In my Ph.D. thesis, I described a large scale RTPCR and cloning pipeline that I developed to obtain several thousand fulllength mammalian (mainly human and mouse) cDNA clones for the NHGRI/NCI Mammalian Gene Collection effort. At that time, the layers of the mammalian transcriptome complexity were not yet fully conceived. In this project, I was deeply involved in strategic and conceptual issues in characterizing transcriptome complexity from different levels. Using the RTPCR pipeline combined with gene prediction algorithms that utilize comparative genomic information, we successfully identified and experimentally verified novel mammalian genes such as fulllength rat homologs of human disease genes. Furthermore, I studied transcriptome complexity including SNPs and alternative splicing in leukemia samples using quantitative PCR, and we developed a trace analysis technology to identify multiple alternative spliced transcripts in one single sequencing reaction. My results suggested that alternatively spliced transcripts show cancer type specific up or down regulation, and the ratio of alternative spliced isoforms may underlie the cancer mechanism.
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birth defects, or other reproductive harm.
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