HO-18 - BIO 320 lectures 18-19 Reading: Text chapter 9,...

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BIO 320 lectures 18-19 Reading: Text chapter 9, pp325-335; ch 10; ch 11.1-11.3 Problems: QB-C Genetictechnology II DNA polymerase and synthetic oligonucleotide primers can be used to label , sequence and amplify defined DNA segments. Nucleotides with a substituted radioactive atom or an added fluorescent tag can be used to make a labelled DNA strand . When separated from its template, this can be used as probe for hybridization experiments. The polymerase chain reaction (PCR) amplifies defined DNA segments exponentially Chain-terminating dideoxynucleotides are used for DNA sequencing template: primer: OH 5' 5' 3' 3' A G G T C T C A C G A T G T C C A G A The basic reaction: single-stranded template , oligonucleotide primer , all 4 nucleotides , DNA polymerase OH 5' 5' 3' 3' A G G T C T C A C G A T G T C C A G A G C G C T A C HO OH 3' T OH 3' A OH 3' G OH 3' C nucleotides polymerase Analysinggenes,genotypesandgenomes . OH 3' T OH 3' C OH 3' G A OH 3' α - 32 P-dATP OH 5' 3' T C C A G A G C G C T A C polymerase H H 3' 2' P P P BASE OH H 3' 2' BASE P P P dideoxynucleotide (ddNTP): no 3' -OH deoxynucleotide (dNTP) C 5' T C C A G A G C template: specific primer: OH 5' 5' 3' 3' A G G T C T C A C G A T G T C C A G A OH 3' T OH 3' A OH 3' G OH 3' C all 4 nucleotides. .. A DNA sequencing reaction contains:
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HO-18 - BIO 320 lectures 18-19 Reading: Text chapter 9,...

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