1 Materials & Methods 4

1 Materials & Methods 4 - 2006 14 Pyrosequencing...

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1 Molecular Biology Sequencing & Synthesis
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3 Fred Sanger Nobel Prize for inventing protein sequencing– demonstrates that proteins are linear chains of amino acids Second Nobel Prize for inventing DNA sequencing
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4 “Sanger sequencing” also called “dideoxy” or “enzymatic” sequencing
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5 Flow chart for Sanger sequencing
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6 Picture of autoradiograph from dideoxy sequencing gel
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7 Enzymatic sequencing using fluorescent labels— a different color for each nucleotide— all can be run in the same lane Migration
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12 Venter et al. 2001 Science > 100 authors!
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13 Science 2006 X-Prize: $10 million Science 2005 Science
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Unformatted text preview: 2006 14 Pyrosequencing: detects pyrophosphate (P-P) released from polymerization 15 16 17 18 19 Pyrosequencing set-up steps: Whole DNA short (300-500 bp) dsDNA sonicate short ssDNA denature attach to bead copy by PCR on bead many copies of same DNA on bead distribute 1 bead into each well pyrosequencing reactions in wells produces light detected by CCD camera add dNTPs one at a time new DNA strand grows when correct dNTP is added 20 Pyrosequencing plate measures 70 x 75 mm, has 1.6 million wells Each DNA strand can be sequenced for ca. 100 bases 21...
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This note was uploaded on 01/21/2012 for the course PDBIO 305 taught by Professor Woods,a during the Fall '08 term at BYU.

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1 Materials & Methods 4 - 2006 14 Pyrosequencing...

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