Exam_II_Summary_Team_4___CE_article - Kingsley Odigie and...

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Kingsley Odigie and Calvin Lee Team 4 Exam II Summary 23 pairs of chromosomes 3 billion base pairs---1/(1/4) 3,000,000,000 possible combinations DNA analysis by RFLP—long process, requires high quality and quantity of DNA Kary Mullis invented PCR STR and other genetic variabilities: Give reproducible results from day to day; resolution of a single base over the range of analysis; precision under 0.17 base pair for size separation; stability over time; insensitivity to matrix effects; relative accuracy though not absolute Methods of determination of variability: probe hybridization; charge based mobility & separation—gel & capillary electrophoresis; partition & ion exchange—HPLC; Conformation—SSCP, heteroduplex polymorphism; size measurement—mass spectrometry Variation in the methods: Probe based methods—can be difficult to detect length variation; HPLC—lacks resolution; MS—has trouble with sizes above 90 bp; Conformational polymorphisms—will not always vary sufficiently; Issue: although PCR is rapid & efficient, sample loads are increasing---untested rape kits nationwide is between 180,000 to 500,000 PCR—Application—has improved speed & detection capabilities; automated system needed to increase sample throughput Capillary Electrophoresis Benefits of capillary electrophoresis: injection, separation and detection are automated; rapid separations are possible and peak information is automatically stored for easy retrieval Electrophoresis process: injection; separation and detection Charges of DNA vary with size
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This note was uploaded on 09/08/2010 for the course JS 113 at San Jose State.

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Exam_II_Summary_Team_4___CE_article - Kingsley Odigie and...

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