9 - Kuo 580.221 30 Sept 2004 Notes for 2005 Molecules Cells...

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Kuo, 580.221 30 Sept 2004 1 Molecules & Cells BME 580.221 Recombinant DNA Techniques Lecture #09, 30 Sept 2004 Scot C. Kuo 30 Sept 2004 Kuo, 580.221 Notes for 2005 • Consider swapping order of presentation for enzymatic amplification before biological amplification – Consider new images from Garland website • How to make storyline less lecture-driven? Ask students to propose strategies to solve technological issues? 30 Sept 2004 Kuo, 580.221 Science vs. Tools can brute-force excellent Efficiency: high stringency moderate Purity: can be absent very stringent Regulation: moderate demands high demands Specificity: few (sequential?) many , simultaneous Reagents: same as Biology column, but circuits must be well-characterized . - what’s modular? - substitutions? few (sequential?); Simplified best many , simultaneous Reactions: Organism Test Tube Tools/Engineering/BioTech Biology 30 Sept 2004 Kuo, 580.221 Improving Specificity (discrimination) • Strategies: – Limit number of reagents/choices (partially purified system) – Control association/affinities by altering temperature, pH, ionic strength (salt), cofactors – Mix reagents from different organisms (e.g. viruses or thermophilic bacteria) – Use reagents from artificial sources 30 Sept 2004 Kuo, 580.221 Biological synthesis of NA RNA subunits: rNTP (rATP, rGTP, rCTP, rUTP) DNA subunits: dNTP (dATP, dGTP, dCTP, dTTP) Example of exploiting natural process as a tool? 30 Sept 2004 Kuo, 580.221 Automated Sanger Sequencing How to make primer? Why is it specific?
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This note was uploaded on 04/15/2008 for the course BME 580.221 taught by Professor Dr.vidal during the Fall '08 term at Johns Hopkins.

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9 - Kuo 580.221 30 Sept 2004 Notes for 2005 Molecules Cells...

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