Why bother with a new technology

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Why bother with a new technology?
Current (2005) 1.00 per kilobase 20 bp /
second 100 Gb / year
NHGRI Goals ? 100x, 5yrs ? 10,000x, 15yrs
Collins, Morgan, Patrinos (2003)
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Growth of GenBank
60
50
40
30
Base pairs of DNA (billions)
Sequences (millions)
20
10
0
1985
2000
1995
1990
December 1982
September 2005
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Completed Genome Sequences (roughly)
And growing
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gt100,000 species are represented in GenBank
all species 128,941 viruses 6,137 bacteria 31,
262 archaea 2,100 eukaryota 87,147
Table 2-1 Page 17
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Absolutely requires a correctly paired nucleotide
so as not to allow errors This requirement means
cant elongate if there is NO nucleotide either -
thus a primer is necessary
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Genome Sequencing Visit http//jgi.doe.gov/edu
cation/how/index.html for more explanation
Hydroshearing
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Insertion of fragments into bacterial plasmids
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Transformation of plasmids into bacteria via
electroporation
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Growth of colonies on an agar plate
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Robotic colony picking and distribution to 384
well plate (one colony per well)
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Lysis of bacterial cultures at 95 C
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Rolling circle amplification of plasmid DNA Use
phi29 polymerase
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Transfer amplified DNA to new plate for cycle
sequencing
Add cycle sequencing reagents
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Purify DNA from sequencing reaction with magnetic
beads
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Bar coding and capillary electrophoresis and
laser scanning
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Computational assembly of fragments into contigs
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Quality control
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