ch09 - 3/22/2011 DNA-Based Technologies I) DNA Technologies...

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3/22/2011 1 I) DNA Technologies II) Protein Technologies DNA-Based Technologies Eschericia coli is a biochemist’s best friend DNA Cloning Here, it enables the cloning and amplification of a eukaryotic DNA fragment of interest Genetically tractable ~20 minute doubling time Grows in media and on agar plates
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3/22/2011 2 Some Enzymes Used in Molecular Biology You will find these enzymes in practically every biochemistry lab Taq polymerase replicates DNA in PCR reactions Common Restriction Enzymes Notice palindromic sequences; often the restriction enzymes are dimeric “Sticky ends” or “blunt ends” are created by cleavage Can estimate frequency of site in DNA: for 50% GC content, 1 EcoRI site every 4 6 = 4,096 bases
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3 Inserting genomic DNA fragments into self replicating plasmids using restriction enzymes and ligase These plasmids need an origin of replication so they can multiply in the host cell. They generally have a high copy number. Polylinkers
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ch09 - 3/22/2011 DNA-Based Technologies I) DNA Technologies...

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