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The Polymerase Chain Reaction. The small sample of DNA serves as template for DNA polymerase Make complementary primers Add primers in more than 1000-fold excess Heat to make ssDNA, then cool Run DNA polymerase (usually Taq) Repeat heating, cooling, polymerase cycle. - PowerPoint PPT Presentation
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• The small sample of DNA serves as template for DNA polymerase
• Make complementary primers • Add primers in more than 1000-fold excess • Heat to make ssDNA, then cool • Run DNA polymerase (usually Taq) • Repeat heating, cooling, polymerase cycle
The Polymerase Chain Reaction
The use of PCR in forensic science
POLYMERASE CHAIN
REACTION
• After cleavage of a plasmid (cloning vector) with a restriction enzyme, a foreign DNA fragment can be inserted
• Ends of the plasmid/fragment are closed to form a "recombinant plasmid"
• Plasmid can replicate when placed in a suitable bacterial host
DNA Cloning
Genomic DNA library & cDNA library
Production of large amounts of a protein by cloning the protein-coding DNA sequence (gene) in a plasmid expression vector
DNA Chips
What are the challenges?
• Error: Molecular operations are not perfect.• Reversible and Irreversible Error• Efficiency: How many molecules
contribute?• Encoding problem in molecules is difficult• Scaling to larger problems
What are the challenges for Computer Science?
• Discover problems DNA Computers are good at– Messy reactions as positive– Evolvable, not programmable
• Characterize complexity for DNA computations with bounded resources
• New notions of what a “computation” is?
What are the challenges for molecular biology?
• Develop computation-specific protocols• Better understanding of basic mechanisms
and properties• Better characterization of processes• Measures of reliability and efficiency• Advanced understanding of biomolecules
other than DNA and RNA
What developments can we expect in the near-term?
• Increased use of molecules other than DNA• Evolutionary approaches• Continued impact by advances in molecular
biology• Some impact on molecular biology by DNA
computation• Increased error avoidance and detection
What are the long-term prospects?
• Cross-fertilization among evolutionary computing, DNA computing, molecular biology, and computation biology
• Niche uses of DNA computers for problems that are difficult for electronic computers
• Increased movement into exploring the connection between life and computation?
Where can I learn more?• Web Sites:
• http://www.wi.leidenuniv.nl/~jdassen/dna.html• http://dope.caltech.edu/winfree/DNA.html• http://www.msci.memphis.edu/~garzonm/bmc.html• (Conrad) http://www.cs.wayne.edu/biolab/index.html
• DIMACS Proceedings: DNA Based Computers I (#27), II (#44), III (#48), IV (Special Issue of Biosystems), V (MIT, June 1999)• Other: Genetic Programming 1 (Stanford, 1997), Genetic Programming 2 (Wisconsin-Madison, 1998), IEEE International Conference on Evolutionary Computation (Indianapolis, 1997)• G. Paun (ed.), Computing with Biomolecules: Theory and Experiment, Springer-Verlag, Singapore 1998.• “DNA Computing: A Review,” Fundamenta Informaticae, 35, 231-245.