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Lecture Handout 10 - BIOT 101 Lecture Handout 9 DNA...

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BIOT 101 Lecture Handout 9 DNA Sequencing and the Human Genome Project Vocabulary dideoxynucleotide sequencing chain termination sequencing Fred Sanger dideoxynucleotides polyacrylamide polyacrylamide gel electrophoresis (PAGE) autoradiography Objectives Be able to list and explain the benefits of know the DNA sequence of an organism. Be able to explain how dideoxynucleotides are used in the Sanger method of DNA sequencing. Understand why polyacrylamide electrophoresis is important in this process. Understand how fluorescent tags are used in DNA sequencing. Be able to interpret a “tradition” sequencing gel, a fluorescence-tagged sequencing gel and the graphic representation from a high-throughput sequencing run. Read Genomics and its impact on Science and Society . 1
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BIOT 101 Lecture Handout 9 DNA Sequencing and the Human Genome Project I. DNA sequencing is the process of determining the exact order of the bases of a length of DNA. The most commonly used method of sequencing is referred to as dideoxy sequencing or chain termination sequencing . It is also referred to as the “Sanger method” because it was developed by Fred Sanger in 1977 (for which he won his second Nobel prize). Modifications of this method have allowed amazingly rapid accumulation of sequence data. This data has many significant uses. A. Generation of sequence data allows for the prediction of amino acid sequence of proteins coded for by genes. With increased computational capabilities, this will eventually allow molecular biologists to predict the shape of proteins without the need to analyze the protein directly (a very tedious task). This will have profound effects on controlling the actions of proteins and predicting their relationships with other proteins. B. Identification of regulatory regions of DNA that are involved in the control of gene expression. This has allowed for greater understanding of gene expression and will lead to increased ability to control the level of expression of specific genes in vivo for both normal and genetically modified organisms.
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