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Text Notes BIO130 Lab 2

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University of Toronto St. George
Jane Mitchell

Lab 2 Text Notes pg 532534 540541 534Analyzing and Manipulating Datacleavage of DNA at specific sites by restriction nucleasesallows for isolation and manipulation of individual genesDNA ligationdesign and construct DNA molecules not in natureDNA cloninguse of cloning vectors or polymerase chain reaction portion of DNA repeatedly copiedgenerates billions of identical moleculesnucleic acid hybridizationcan find specific DNARNA sequence of basis of its ability to selectively bind complementary nucleic acid sequencesdetermine sequence of DNA nucleotidesidentify genes deduce amino acid sequence of proteins they encodemonitor levels of mRNA produced by every gene in a cell with nucleic acid arraystens of thousands of hybridization reactions take place simultaneouslyRestriction Nucleasesgenenot a discrete entitysmall region of much longer DNA moleculeDNA molecules cannot be easily separated because they consist of approximately the same mixture of the same four nucleotides on the basis of different charges and binding propertiesrestriction nucleasesenzymes purified from bacteriacleave long strands of DNA double helices at specific site into fragments of defined sizesrestriction nucleases each have different sequence specificitiesan enzyme can create a DNA fragment
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