g640Lecture12B

g640Lecture12B - nucleic acid variation indirect DNA...

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nucleic acid variation indirect – DNA/DNA hybridization – enzymatic degradation restriction enzyme digestion (RFLP) – amplification polymorphism (RAPD) – repeat sequence variation (VNTR / STR) direct sequencing DNA POLYMORPHISMS DIRECT SEQUENCE DETERMINATION – CLONING – ORGANELLE GENOMES – PCR AMPLIFICATION OF TARGET GENES / SEQUENCING GENE STRUCTURE 5’ Flanking 3’ Flanking Promoter start ---------------------------------stop-----5’untranslated 5’untranslated DNA SEQUENCES Sequencing procedures developed by – Sanger - dideoxy chain termination – Maxam & Gilbert - chemical termination Most sequencing now performed by dideoxy sequencing DIDEOXY NUCLEOTIDES ATCGGGCTATC
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DIDEOXY NUCLEOTIDES ATCGGGCTATC A* ....................... AT* ..................... ATC* ................... ATCG* ................ ATCGG* ............. ATCGGG* ........... ATCGGGC* ........ ATCGGGCT* ...... ATCGGGCTA* DIDEOXY NUCLEOTIDES ATCGGGCTATC A* ....................... ATCGGGCTA* .... DIDEOXY NUCLEOTIDES ATCGGGCTATC AT* ..................... ATCGGGCT* ...... ATCGGGCTAT* .. DIDEOXY NUCLEOTIDES ATCGGGCTATC ATC* ................... ATCGGGC* ........ ATCGGGCTATC* DIDEOXY NUCLEOTIDES ATCGGGCTATC ATCG* ................
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g640Lecture12B - nucleic acid variation indirect DNA...

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