DNA sequencing

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ATGTTCTATCCCATTCATTTTGACGTTATTGTTGTTGGAGGAGGTCATG
CTGGGACAGAAGCAGCTTTGGCTTCTGCTAGGATGCAATGTAATACGCTT
TTGATTACTCATAATATTGATACTCTTGGACAGATGTCTTGTAATCCGGC
CGTTGGTGGCATAGGGAAAGGACATTTAGTGAAAGAAATTGATGCGATGG
GTGGATCGATGGCTTATGCTATCGATCAATCAGGAATTCAATTTAGAGTA
CTTAATAGTAGCAAAGGAGCTGCTGTTAGAGCAACACGTGCTCAGGCAGA
TAAAATATTATATCGTCAAGCAATACGTAGTATTCTTGAATATCAAAAAT
TTTTGTTGGTTATTCAAGCGTCAGTAGAAGATTTAATAGTTAGTGGGAAC
AAGATTGTTGGAGTAATTACTCCAAAATTAGGAATGAAATTTAGTGGTAC
GTCTGTTGTGTTGACAACCGGAACCTTTCTCAATGGTAAGATTCATATTG
GGATGAATAATTTTAGAGGAGGTCGATCTGGAGATTCGGAGTCATCGTCA
TTGTTATCAGAGCGATTGAAAGAATTGTCTTTTCAGATTAGTCGGTTAAA
AACGGGTACTTCTCCTCGTGTGCATACCAAAGGGATAAATTTTGGTTCTT
TACGTGCACAATATAGTGATGATCCCATTCCTGTGTTTTCATTTATAGGA
TCTACAAAACTACATCCTACGCAGGTGCCTTGTTATATTACCCATACTAA
TAACAAAACACATGAAATAGTTAGATCAAATTTATATCAGAGTCCTATGT
ATACAGGTTTAATAAAAGGAATAGCACCTCGGTATTGTCCATCTATAGAA
GACAAAATAACTCGTTTTTCAGATCGTAATGCTCATCAAATTTTTTTAGA
ACCTGAAGGTTTGACAACACCTGAAGTATATCTTAATGGTATTTCTACTA
GCTTACCTTTTTGTGTACAAATGCAAATGATTAAATCAATTCAGGGATTA
GAGAATGCTTGTATAATTAGGCCAGGTTATGCTATTGAATACGATTTTTT
TGATCCTCGTGATTTAAAATTAACATTAGAGAGTAAAATTATTTCTGGAT
TATTTTTTTCTGGTCAAATTAATGGTACTACTGGTTATGAAGAAGCAGCT
GCCCAAGGATTATTGGCCGGAATTAATGCAGCTAGGTTTTCCAAAAATAA
AGAAGGATGGTACCCTAGAAGAGATCAGGCATATTTAGGGGTGCTTGTAG
ATGATCTCTGTACACATGGAACAGAAGAACCTTATCGTATGTTTACTTCG
CGTGCTGAATATCGTCTGTCTTTACGTGAGGATAATGCTGATTTAAGATT
AACTGAAATCGCGCGACAGTTAGGTTTGATAGATGAGTCACGGTGGAAAG
CTTTTTGTTGCAAAAAAGAAAACATTGAAAAAGAACGTCAGAGATTACGT
AATACTTATATTTTCCCGTATAGCTCAGATGTTGCACAATTAAATAATTT
TCTTAAAACACCTTTAACACATGAAACAAATGGCGAAGATCTTTTGAGAA
GGCCAGAAATTAATTATAAAAAGTTATCTCAACTAAGTACTTTTAGTCCA
TCTATATTGGATCGTCAAGTATTCGAGCAAATTGAAATTCAAATAAAATA
TGAAGGTTATATTCGTCATCAACAAGAAGAAATAAAGAGACATATTTATA
ATGAAAATACATTGTTACCGACTGATATAGATTTCAATATTGTTTCTGGA
TTATCTCAGGAGGTCATTGACAAACTCAATAATTATAAACCTTATTCTAT
TGGGCAGGCTTCTCGCATTTCTGGTATCACTCCTGCGGCTATTTCTAATT
TATTGGTTTGGTTAAAAAAACAAGGTTTGTTAGAGCATAATACATGCTAA
TCTGTTGAATAATGTATTTTCTATTAATCAAGGTATTATTTAATTTCCTA
TATAGGGTGTTTGTGATTGTTACAATATGGAGAGAGTTGATCGTGTAGAA
AACCGGTATATTTATGTCTTTTCCAATAATAATTTTATAGAAATAATAAT
AGTAATTAGATTATTATTTTAATGGGTTTGTTATGTATGGCATTATTTTT
AATATTATACGGTTGTTTATACGTATCTAAATTTGCGCTTTGAATTTTTT
TGAAAATAGGATTATACTGAACTTTTAGTTGTATACAAGTGTATTATTTC
ATATTTTTGAATAAATATACGTGCTGTACTAATGGTTATTAATTACAAGA
TAAATTACTTGTCGACCGAACTATAGTTCGGTCGTGTGTTTGTTTCTGTA
AGTTGTAACATATTATGTGAAATTTACTAATTAATTCATAGTGTCATTTT
AATTCTCTTTTATTGCATTAGTATATGGTTGTGTAAAAAGAGAGAGAGTG
TTCTTGTTATTTTCTATTAGTACTAATTTTTTAAAAGTTGATGCAGGCAC
TGTCGTTCTTTTCGGATACAACATCATTGTAAGTTATATATAAAAATGAT
TTAGAATTTCGACATGTTAGTGACAAGTTGTATATCGAGGTTGTAATGTT
GTATTATACAAGAACATTGTAAAATATTTATAGTATATTGCAGCCAATAT
TTTTTGAAAGGTGGTTTATGTCAGGAATCAAAGGCACTTCCCAAGAATAT
ATTGGACATCATTTATATCATCTGCAATTTGATTTAAGTACTTTTTCGTT
AGTGAGCTCGGAGAATACTTCTTCATTCTGGGTATTAAATGTAGATTCGA
TGTTTTTTTCAATATTATTGGCTACTTTATTTTTATTAATTTTCGGTCGT
TTGGCTACAGTGGCAACTTATGCGGTTCCCACAAAACTGCAAGTGTTTAT
TGAGTTAGTAATATTGTTTATTGATAGCAATGTAAAAGATATGTTTCATG
GTAAAAATAAACTAATTGCGCCATTATCTATGACGGTTTTTGTTTGGATT
TTTTTAATGAATACCATGGATTTATTTCCCATAGATTTATTTCCTGCTAT
AGCTAAATTGTTAGGATTACCTGCTTTACGTGTTGTGCCGTCTGCCGATG
TGAATATCACTTCTTCGCTAGCTTTAAATGTATTTGTACTTGTTATGTAC
TACAATATTTACGTTAATGGCGTTCATGGTTTTATTAAAGGACTGATGTA
TCATCCATTTAATCATCCAACATGTATTCCTATTAATTTTATTATTGAAG
TTGTTAGTTTGTTATCTAAACCAGTATCACTTAGTCTTAGATTATTTGGC
AATATGTATTCTGGTGAGTTGATTTTTATTTTGATATCTGGTTTATTACC
ATGGTGGGGACAATGGGTTTTAAATTTACCATGGGCTATTTTTCATATTT
TAGTCGTTACATTACAGGCTTTTATTTTTATGGTTTTAACGGTGATTTAT
TTATCTACAGCCCATGACTCCTGTTAAAATGAATCATGTAATACAGAAGA
TTGCAGAGAGGTTATTATGGAACATTTAAATTTTGATATGTTATATATTG
CTGCAGCAATAATGATGGGATTAGCAGCAATTGGAGCGGCTATCGGTATT
GGCATCTTAGGTAGTAAATTTTTAGAGGGGGCTGCACGTCAACCAGATCT
CATTCCGATCCTTCGAACTCAGTTTTTTATTGTTATGGGATTGGTTGATG
CAATACCAATGATTACTGTAGGTCTTGGTTTGTATGTGATGTTTTCTGCG
GTTTAACAATGAAAATTAATTGCATGTATTTATTATATTGATTATTAAAT
ATCAGGATTTGCGCTGTGAATCTTAATGCAACAATATTAGGTCAAACTAT
TTCATTTGTTTTGTTTGTTTGGTTTTGTATGAAGTATGTGTGGTATCCAT
TTATATCTATTATTGAGAAACGCCAAAAAGAAATTTCTGATAATTTAGTT
TCTGCTACTCATGCCAAAACAGAATCTGAGCGTGTCAATGCTGAAGCTTT
GCTTTGTTTGAGACAGGCTCGGGTCAAAGCTCAAGAAATTATAAAACAAG
CAAATAAATGTAAAATGCAAATAATTAATGAAGCTAAACATGAAGCTGAA
AAGGAGCAAAGTAGAATTTTATCTCAAGCGCGAGAACAGATTATTTATGA
AAGAAAACGTGTTACTGACGAATTAAGAAAGCAAATTAGCGAACTTGTAA
TTGAGGGTACAGAGAAAGTTATAGAACATTCTATAAATGAAATGATTGAT
ATAGATCTTTTAAATAATATTATTAATACGTTGTCATATAAGGATTAGAT
GTCTAGTATGCTTGTGGTTGCGCGTACGTATGCTCAAGCTATATTTGATA
TAGCTGTAGAACAGAAAAATATAAACAAGTGGAAATCAGTTCTTGATTTA
TTTTCTGAGATTAGTCTAAATAGACTAGTACAATCTTTATTTTTTAGATG
TTTAGAACCAAAAAGATTGTCAGATATATTTATTGCTATTTGTGAAGATT
ACCAAAAGAAACAAGTTGATACCTTCAGTAAAAATATAATCTATATTATG
GCGGAAAATAATCGTTTATTATTATTACCAATTGTATTTAAAGAGTTTAC
TTATTTATGTTCTATATATGTTCATACTGTAGAAATAGAAATTATCTCTG
CTTGGCCTTTGAAGTATAATCAGCTGAAAAAAATTACTGATATAATGGCT
AAACGTTTATCTAAAACAGTGAATCCAGTACACAAAGTAGATAAAGATAT
ATTGGCTGGTGTAATTATTCGTATTGGAGATACTGTGATTGATGGAAGTA
TACGTGGACGTATTTTTCGCTTAAATCACGTACTACAATCTTAATATTTT
AACAGTAGTTAAAAGGTATAAACAAAAGATTATGCAATTAAATTCAAATG
AAATTTCAGATAATGGAGAAGTTATTATTAATGATTTAAAGTTATTTTAT
AATAAAGCTAGACAAACTAAAATTACAGAAGAACTTACAGAAATTGTTTC
AGGAGCTTCTGTAATATAAACTTAAAAGATAGTTAGAGGTATAATTAATA
TGAGTTCTGGAAAAATTGTCCAGGTTATTGGAGCGGTGGTTGATGTTGCG
TTCAATCAAGATGTGGTACCGACTGTATACCATGCACTTGAGGTGTAATA
CTTATATTTTCCCGTATAGCTCAGATGTTGCACAATTAAATAATTTTCTT
AAAACACCTTTAACACATGAAACAAATGGCGAAGATCTTTTGAGAAGGCC
AGAAATTAATTATAAAAAGTTATCTCAACTAAGTACTTTTAGTCCATCTA
TATTGGATCGTCAAGTATTCGAGCAAATTGAAATTCAAATAAAATATGAA
GGTTATATTCGTCATCAACAAGAAGAAATAAAGAGACATATTTATAATGA
AAATACATTGTTACCGACTGATATAGATTTCAATATTGTTTCTGGATTAT
CTCAGGAGGTCATTGACAAACTCAATAATTATAAACCTTATTCTATTGGG
CAGGCTTCTCGCATTTCTGGTATCACTCCTGCGGCTATTTCTAATTTATT
GGTTTGGTTAAAAAAACAAGGTTTGTTAGAGCATAATACATGCTAATCTG
TTGAATAATGTATTTTCTATTAATCAAGGTATTATTTAATTTCCTATATA
GGGTGTTTGTGATTGTTACAATATGGAGAGAGTTGATCGTGTAGAAAACC
GGTATATTTATGTCTTTTCCAATAATAATTTTATAGAAATAATAATAGTA
ATTAGATTATTATTTTAATGGGTTTGTTATGTATGGCATTATTTTTAATA
TTATACGGTTGTTTATACGTATCTAAATTTGCGCTTTGAATTTTTTTGAA
AATAGGATTATACTGAACTTTTAGTTGTATACAAGTGTATTATTTCATAT
TTTTGAATAAATATACGTGCTGTACTAATGGTTATTAATTACAAGATAAA
TTACTTGTCGACCGAACTATAGTTCGGTCGTGTGTTTGTTTCTGTAAGTT
GTAACATATTATGTGAAATTTACTAATTAATTCATAGTGTCATTTTAATT
CTCTTTTATTGCATTAGTATATGGTTGTGTAAAAAGAGAGAGAGTGTTCT
TGTTATTTTCTATTAGTACTAATTTTTTAAAAGTTGATGCAGGCACTGTC
GTTCTTTTCGGATACAACATCATTGTAAGTTATATATAAAAATGATTTAG
AATTTCGACATGTTAGTGACAAGTTGTATATCGAGGTTGTAATGTTGTAT
TATACAAGAACATTGTAAAATATTTATAGTATATTGCAGCCAATATTTTT
TGAAAGGTGGGTGGATCGATGGCTTATGCTATCGATCAATCAGGAATTCA
ATTTAGAGTACTTAATAGTAGCAAAGGAGCTGCTGTTAGAGCAACACGTG
CTCAGGCAGATAAAATATTATATCGTCAAGCAATACGTAGTATTCTTGAA
TATCAAAAATTTTTGTTGGTTATTCAAGCGTCAGTAGAAGATTTAATAGT
TAGTGGGAACAAGATTGTTGGAGTAATTACTCCAAAATTAGGAATGAAAT
TTAGTGGTACGTCTGTTGTGTTGACAACCGGAACCTTTCTCAATGGTAAG
ATTCATATTGGGATGAATAATTTTAGAGGAGGTCGATCTGGAGATTCGGA
GTCATCGTCATTGTTATCAGAGCGATTGAAAGAATTGTCTTTTCAGATTA
GTCGGTTAAAAACGGGTACTTCTCCTCGTGTGCATACCAAAGGGATAAAT
TTTGGTTCTTTACGTGCACAATATAGTGATGATCCCATTCCTGTGTTTTC
ATTTATAGGATCTACAAAACTACATCCTACGCAGGTGCCT
DNA sequencing Why? Identifies Organisms How?
Dideoxy Sequencing Introduces Biological
Machines Where? Universities, MBL Societal
Impact? Forensics, Insurance
DNA detective
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Review of Project
PCR
DNA extraction
Insect identification
?
DNA sequencing
Gel electrophoresis
ACAGATGTCTTGTAATCCGGCCGTTGGTGGCATAGGGAAAGGACATTTAG
TGAAAGAAATTGATGCGATGGGTGGATCGATGGCTTATGCTATCGATCAA
TCAGGAATTCAATTTAGAGTACTTAATAGTAGCAAAGGAGCTGCTGTTAG
AGCAACACGTGCTCAGGCAGATAAAATATTATATCGTCAAGCAATACGTA
GTATTCTTGAATATCAAAAATTTTTGTTGGTTATTCA
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How is this organism related to other species?
K
?
C
A
H
F
I
J
D
B
E
G
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• DNA sequences provide characters that are
• Numerous, discrete characters (A, T, C, G)
• Directly comparable across species
• Unlikely to change due to culture conditions
• Can be sampled without ever culturing (important
  for endosymbionts!)

• Molecular phylogenetics
• Inference of evolutionary relationships based on
  molecular data

ACAGATGTCTTGTAATCCGGCCGTTGGTGGCATAGGGAAAGGACATTTAG
TGAAAGAAATTGATGCGATGGGTGGATCGATGGCTTATGCTATCGATCAA
TCAGGAATTCAATTTAGAGTACTTAATAGTAGCAAAGGAGCTGCTGTTAG
AGCAACACGTGCTCAGGCAGATAAAATATTATATCGTCAAGACAGATGTC
TTGTAATCCGGCCGTTGGTGGCATAGGGAAAGGACATTTAGTGAAAGAAA
TTGATGCGATGACAGATGTCTTGTAATCCGGCCGTTGGTGGCATAGGGAA
AGGACATTTAGTGAAAGAAATTGATGCGATGGGTGGATCGATGGCTTATG
CTATCGATCAATCAGGAATTCAATTTAGAGTACTTAATAGTAGCAAAGGA
GCTGCTGTTAGAGCAACACGTGCTCAGGCAGATAAAATATTATATCGTCA
AGCAATACGTGGTGGATCGATGGCTTATGCTATCGATCAATCAGGAATTC
AATTTAGAGTACTTAATAGTAGCAAAGGAGCTGCTGTTAGAGCAACACGT
GCTCAGGCAGATAAAATATTATATCGTCAAGCAATACGTACAGATGTCTT
GTAATCCGGCCGTTGGTGGCATAGGGAAAGGACATTTAGTGAAAGAAATT
GATGCGATGGGTGGATCGATGGCTTATGCTATCGATCAATCAGGAATTCA
ATTTAGAGTACTTAATAGTAGCAAAGGAGCTGCTGTTAGAGCAACACGTG
CTCAGGCAGATAAAATATTATATCGTCAAGCAATACGTCAATACGCGTGC
TCAGGCAGATAAAATATTA
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Molecular Phylogenetics Step 1. Select a DNA
region that is homologous, or similar across
species due to common ancestry.
Secondary structure of 16S rRNA in E. coli
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Importance of Ribosomes in Cell Biology

• Ribosomes are part of the machinery that
  synthesize cell proteins
• Ribosomes are so important that they are
  ubiquitous and highly conserved

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• 2. Amplify and Sequence this region across
  isolates.

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3. Estimate relationships based on extent of DNA
similarity.
At variable DNA positions, related groups will
tend to share the same nucleotide. The sheer
number of characters is helpful to distinguish
the phylogenetic signal from noise.
ATGTTGGCAGTCCGATGTAAGC
ATGTTGGCAGTCCGATGTAAGC
ATGTTGGCAGTCCGATGTAACC
ACGGTAGCAGTCTGATGTATCC
ACGGTAGCAGTCTGATGTATCC
ACGGTAGCAGTCTGATGTATCC
CTGCTGGTAGTCGTTTGTAACC
CTGCTGGTAGTCGTTTGTAACC
CTGCTGGCAGTCGGTTGTAACC
ATGCTGGCAGTCGGGTGTAACC
Molecular phylogeny of taxa A-I.
ATGGTGGCAGTCGGGTGTCACC
Colored letters different from top sequence
(taxon G)
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The elegant idea behind DNA sequencing
Technology changes quickly, but for many years
weve used Sangers cool trick.
In the 1970s, Sangers group discovered a
fundamentally new method of 'reading' the linear
DNA sequence using special bases called chain
terminators or dideoxynucleotides. This method
is still in use today. What is the basis of
Sangers method?
Shared with Walter Gilbert and Paul Berg
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Fascinating. Now how do we actually sequence ??
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DNA Polymerase Action
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Mechanism of DNA polymerization
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Dideoxy Nucleotide
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Dideoxies block elongation
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DNA Sequencing Reactions
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Gel Electrophoretic Fractionationof Products
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Fractionating DNA Molecules

• DNA is negatively charged at neutral pH
• DNA will travel towards a positive charge
• DNA electrophoresed in a gel will travel
  according to its chain length

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Detecting DNA

• Early experiments used radio-isotopically labeled
  DNA (primers)
• Now we use fluorescently labeled
  dideoxynucleotides

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Perform cycle sequencing in PCR machine
Reaction steps seems a lot like PCR, but not
quite
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Use of capillaries filled with a matrix for
higher through-put of sequencing reactions.
e.g. ABI 3730
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In sum From PCR to sequence data
2. Set up sequencing reaction
1. Purify PCR product
3. Perform cycle sequencing
5. Read order of terminators (DNA sequence)
4. Resolve sequence fragments
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• Quality of sequence data may vary, depending on
• Purity and concentration of template DNA
• Presence of extra PCR bands (artifacts)
• Quality of dye-terminators, electrophoresis
  matrix, and other reagents
• Ideally, look at chromatograms and convince
  yourself that base calls are robust.

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DNA sequence assembly Combining sequence reads
to build the entire sequence of the template
DNA

• Sequence reads typically range from 500-800 bp.
  Often the total DNA fragment is much longer.
• Regions of overlap among sequence reads
• Let us piece together the linear puzzle of the
  template DNA sequence.
• Often confirm base calls and improve overall add
  data quality.

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Major sequencing centers have facilities to
sequence multiple full genomes each year

• Due to genomic research over the past decade, DNA
  sequencing has become
• Increasingly automated
• Higher-throughput
• Less per reaction

• And (good news) more accessible to researchers
  and educators

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Possible avenues for DNA sequencing
Many public universities to have DNA sequencing
facilities in house. Such facilities are
usually able to run small numbers of reactions
for a cost of 10-15 per reaction, but often
much less for internal users with ties to the
university. Ideal situation Team up with
faculty at your local university to act as
facilitators.

• Even if DNA sequencing not an option for
  classroom labs, important to discuss
• how DNA sequencing is performed,
• how data is evaluated,
• what information can be obtained from the data,
• what biologically important concepts can be
  learned,
• ethical issues surrounding its interpretation
  and use.

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Ethical issues involving DNA sequencing Truro A
tranquil Cape community shaken by the murder of a
young woman in January 2002
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Should police have requested DNA of every man in
Truro??
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Should scientists attempt to clone the
now-extinct Thylacine to revive this species??
(The technology will be there eventually.)
Thylacine pup preserved in alcohol in 1866. Its
cells could be used for cloning. By chance this
Thylacine was stored in a jar of alcohol rather
than formalin, which would have destroyed the DNA.
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ATGTTCTATCCCATTCATTTTGACGTTATTGTTGTTGGAGGAGGTCATG
CTGGGACAGAAGCAGCTTTGGCTTCTGCTAGGATGCAATGTAATACGCTT
TTGATTACTCATAATATTGATACTCTTGGACAGATGTCTTGTAATCCGGC
CGTTGGTGGCATAGGGAAAGGACATTTAGTGAAAGAAATTGATGCGATGG
GTGGATCGATGGCTTATGCTATCGATCAATCAGGAATTCAATTTAGAGTA
CTTAATAGTAGCAAAGGAGCTGCTGTTAGAGCAACACGTGCTCAGGCAGA
TAAAATATTATATCGTCAAGCAATACGTAGTATTCTTGAATATCAAAAAT
TTTTGTTGGTTATTCAAGCGTCAGTAGAAGATTTAATAGTTAGTGGGAAC
AAGATTGTTGGAGTAATTACTCCAAAATTAGGAATGAAATTTAGTGGTAC
GTCTGTTGTGTTGACAACCGGAACCTTTCTCAATGGTAAGATTCATATTG
GGATGAATAATTTTAGAGGAGGTCGATCTGGAGATTCGGAGTCATCGTCA
TTGTTATCAGAGCGATTGAAAGAATTGTCTTTTCAGATTAGTCGGTTAAA
AACGGGTACTTCTCCTCGTGTGCATACCAAAGGGATAAATTTTGGTTCTT
TACGTGCACAATATAGTGATGATCCCATTCCTGTGTTTTCATTTATAGGA
TCTACAAAACTACATCCTACGCAGGTGCCTTGTTATATTACCCATACTAA
TAACAAAACACATGAAATAGTTAGATCAAATTTATATCAGAGTCCTATGT
ATACAGGTTTAATAAAAGGAATAGCACCTCGGTATTGTCCATCTATAGAA
GACAAAATAACTCGTTTTTCAGATCGTAATGCTCATCAAATTTTTTTAGA
ACCTGAAGGTTTGACAACACCTGAAGTATATCTTAATGGTATTTCTACTA
GCTTACCTTTTTGTGTACAAATGCAAATGATTAAATCAATTCAGGGATTA
GAGAATGCTTGTATAATTAGGCCAGGTTATGCTATTGAATACGATTTTTT
TGATCCTCGTGATTTAAAATTAACATTAGAGAGTAAAATTATTTCTGGAT
TATTTTTTTCTGGTCAAATTAATGGTACTACTGGTTATGAAGAAGCAGCT
GCCCAAGGATTATTGGCCGGAATTAATGCAGCTAGGTTTTCCAAAAATAA
AGAAGGATGGTACCCTAGAAGAGATCAGGCATATTTAGGGGTGCTTGTAG
ATGATCTCTGTACACATGGAACAGAAGAACCTTATCGTATGTTTACTTCG
CGTGCTGAATATCGTCTGTCTTTACGTGAGGATAATGCTGATTTAAGATT
AACTGAAATCGCGCGACAGTTAGGTTTGATAGATGAGTCACGGTGGAAAG
CTTTTTGTTGCAAAAAAGAAAACATTGAAAAAGAACGTCAGAGATTACGT
AATACTTATATTTTCCCGTATAGCTCAGATGTTGCACAATTAAATAATTT
TCTTAAAACACCTTTAACACATGAAACAAATGGCGAAGATCTTTTGAGAA
GGCCAGAAATTAATTATAAAAAGTTATCTCAACTAAGTACTTTTAGTCCA
TCTATATTGGATCGTCAAGTATTCGAGCAAATTGAAATTCAAATAAAATA
TGAAGGTTATATTCGTCATCAACAAGAAGAAATAAAGAGACATATTTATA
ATGAAAATACATTGTTACCGACTGATATAGATTTCAATATTGTTTCTGGA
TTATCTCAGGAGGTCATTGACAAACTCAATAATTATAAACCTTATTCTAT
TGGGCAGGCTTCTCGCATTTCTGGTATCACTCCTGCGGCTATTTCTAATT
TATTGGTTTGGTTAAAAAAACAAGGTTTGTTAGAGCATAATACATGCTAA
TCTGTTGAATAATGTATTTTCTATTAATCAAGGTATTATTTAATTTCCTA
TATAGGGTGTTTGTGATTGTTACAATATGGAGAGAGTTGATCGTGTAGAA
AACCGGTATATTTATGTCTTTTCCAATAATAATTTTATAGAAATAATAAT
AGTAATTAGATTATTATTTTAATGGGTTTGTTATGTATGGCATTATTTTT
AATATTATACGGTTGTTTATACGTATCTAAATTTGCGCTTTGAATTTTTT
TGAAAATAGGATTATACTGAACTTTTAGTTGTATACAAGTGTATTATTTC
ATATTTTTGAATAAATATACGTGCTGTACTAATGGTTATTAATTACAAGA
TAAATTACTTGTCGACCGAACTATAGTTCGGTCGTGTGTTTGTTTCTGTA
AGTTGTAACATATTATGTGAAATTTACTAATTAATTCATAGTGTCATTTT
AATTCTCTTTTATTGCATTAGTATATGGTTGTGTAAAAAGAGAGAGAGTG
TTCTTGTTATTTTCTATTAGTACTAATTTTTTAAAAGTTGATGCAGGCAC
TGTCGTTCTTTTCGGATACAACATCATTGTAAGTTATATATAAAAATGAT
TTAGAATTTCGACATGTTAGTGACAAGTTGTATATCGAGGTTGTAATGTT
GTATTATACAAGAACATTGTAAAATATTTATAGTATATTGCAGCCAATAT
TTTTTGAAAGGTGGTTTATGTCAGGAATCAAAGGCACTTCCCAAGAATAT
ATTGGACATCATTTATATCATCTGCAATTTGATTTAAGTACTTTTTCGTT
AGTGAGCTCGGAGAATACTTCTTCATTCTGGGTATTAAATGTAGATTCGA
TGTTTTTTTCAATATTATTGGCTACTTTATTTTTATTAATTTTCGGTCGT
TTGGCTACAGTGGCAACTTATGCGGTTCCCACAAAACTGCAAGTGTTTAT
TGAGTTAGTAATATTGTTTATTGATAGCAATGTAAAAGATATGTTTCATG
GTAAAAATAAACTAATTGCGCCATTATCTATGACGGTTTTTGTTTGGATT
TTTTTAATGAATACCATGGATTTATTTCCCATAGATTTATTTCCTGCTAT
AGCTAAATTGTTAGGATTACCTGCTTTACGTGTTGTGCCGTCTGCCGATG
TGAATATCACTTCTTCGCTAGCTTTAAATGTATTTGTACTTGTTATGTAC
TACAATATTTACGTTAATGGCGTTCATGGTTTTATTAAAGGACTGATGTA
TCATCCATTTAATCATCCAACATGTATTCCTATTAATTTTATTATTGAAG
TTGTTAGTTTGTTATCTAAACCAGTATCACTTAGTCTTAGATTATTTGGC
AATATGTATTCTGGTGAGTTGATTTTTATTTTGATATCTGGTTTATTACC
ATGGTGGGGACAATGGGTTTTAAATTTACCATGGGCTATTTTTCATATTT
TAGTCGTTACATTACAGGCTTTTATTTTTATGGTTTTAACGGTGATTTAT
TTATCTACAGCCCATGACTCCTGTTAAAATGAATCATGTAATACAGAAGA
TTGCAGAGAGGTTATTATGGAACATTTAAATTTTGATATGTTATATATTG
CTGCAGCAATAATGATGGGATTAGCAGCAATTGGAGCGGCTATCGGTATT
GGCATCTTAGGTAGTAAATTTTTAGAGGGGGCTGCACGTCAACCAGATCT
CATTCCGATCCTTCGAACTCAGTTTTTTATTGTTATGGGATTGGTTGATG
CAATACCAATGATTACTGTAGGTCTTGGTTTGTATGTGATGTTTTCTGCG
GTTTAACAATGAAAATTAATTGCATGTATTTATTATATTGATTATTAAAT
ATCAGGATTTGCGCTGTGAATCTTAATGCAACAATATTAGGTCAAACTAT
TTCATTTGTTTTGTTTGTTTGGTTTTGTATGAAGTATGTGTGGTATCCAT
TTATATCTATTATTGAGAAACGCCAAAAAGAAATTTCTGATAATTTAGTT
TCTGCTACTCATGCCAAAACAGAATCTGAGCGTGTCAATGCTGAAGCTTT
GCTTTGTTTGAGACAGGCTCGGGTCAAAGCTCAAGAAATTATAAAACAAG
CAAATAAATGTAAAATGCAAATAATTAATGAAGCTAAACATGAAGCTGAA
AAGGAGCAAAGTAGAATTTTATCTCAAGCGCGAGAACAGATTATTTATGA
AAGAAAACGTGTTACTGACGAATTAAGAAAGCAAATTAGCGAACTTGTAA
TTGAGGGTACAGAGAAAGTTATAGAACATTCTATAAATGAAATGATTGAT
ATAGATCTTTTAAATAATATTATTAATACGTTGTCATATAAGGATTAGAT
GTCTAGTATGCTTGTGGTTGCGCGTACGTATGCTCAAGCTATATTTGATA
TAGCTGTAGAACAGAAAAATATAAACAAGTGGAAATCAGTTCTTGATTTA
TTTTCTGAGATTAGTCTAAATAGACTAGTACAATCTTTATTTTTTAGATG
TTTAGAACCAAAAAGATTGTCAGATATATTTATTGCTATTTGTGAAGATT
ACCAAAAGAAACAAGTTGATACCTTCAGTAAAAATATAATCTATATTATG
GCGGAAAATAATCGTTTATTATTATTACCAATTGTATTTAAAGAGTTTAC
TTATTTATGTTCTATATATGTTCATACTGTAGAAATAGAAATTATCTCTG
CTTGGCCTTTGAAGTATAATCAGCTGAAAAAAATTACTGATATAATGGCT
AAACGTTTATCTAAAACAGTGAATCCAGTACACAAAGTAGATAAAGATAT
ATTGGCTGGTGTAATTATTCGTATTGGAGATACTGTGATTGATGGAAGTA
TACGTGGACGTATTTTTCGCTTAAATCACGTACTACAATCTTAATATTTT
AACAGTAGTTAAAAGGTATAAACAAAAGATTATGCAATTAAATTCAAATG
AAATTTCAGATAATGGAGAAGTTATTATTAATGATTTAAAGTTATTTTAT
AATAAAGCTAGACAAACTAAAATTACAGAAGAACTTACAGAAATTGTTTC
AGGAGCTTCTGTAATATAAACTTAAAAGATAGTTAGAGGTATAATTAATA
TGAGTTCTGGAAAAATTGTCCAGGTTATTGGAGCGGTGGTTGATGTTGCG
TTCAATCAAGATGTGGTACCGACTGTATACCATGCACTTGAGGTGTAATA
CTTATATTTTCCCGTATAGCTCAGATGTTGCACAATTAAATAATTTTCTT
AAAACACCTTTAACACATGAAACAAATGGCGAAGATCTTTTGAGAAGGCC
AGAAATTAATTATAAAAAGTTATCTCAACTAAGTACTTTTAGTCCATCTA
TATTGGATCGTCAAGTATTCGAGCAAATTGAAATTCAAATAAAATATGAA
GGTTATATTCGTCATCAACAAGAAGAAATAAAGAGACATATTTATAATGA
AAATACATTGTTACCGACTGATATAGATTTCAATATTGTTTCTGGATTAT
CTCAGGAGGTCATTGACAAACTCAATAATTATAAACCTTATTCTATTGGG
CAGGCTTCTCGCATTTCTGGTATCACTCCTGCGGCTATTTCTAATTTATT
GGTTTGGTTAAAAAAACAAGGTTTGTTAGAGCATAATACATGCTAATCTG
TTGAATAATGTATTTTCTATTAATCAAGGTATTATTTAATTTCCTATATA
GGGTGTTTGTGATTGTTACAATATGGAGAGAGTTGATCGTGTAGAAAACC
GGTATATTTATGTCTTTTCCAATAATAATTTTATAGAAATAATAATAGTA
ATTAGATTATTATTTTAATGGGTTTGTTATGTATGGCATTATTTTTAATA
TTATACGGTTGTTTATACGTATCTAAATTTGCGCTTTGAATTTTTTTGAA
AATAGGATTATACTGAACTTTTAGTTGTATACAAGTGTATTATTTCATAT
TTTTGAATAAATATACGTGCTGTACTAATGGTTATTAATTACAAGATAAA
TTACTTGTCGACCGAACTATAGTTCGGTCGTGTGTTTGTTTCTGTAAGTT
GTAACATATTATGTGAAATTTACTAATTAATTCATAGTGTCATTTTAATT
CTCTTTTATTGCATTAGTATATGGTTGTGTAAAAAGAGAGAGAGTGTTCT
TGTTATTTTCTATTAGTACTAATTTTTTAAAAGTTGATGCAGGCACTGTC
GTTCTTTTCGGATACAACATCATTGTAAGTTATATATAAAAATGATTTAG
AATTTCGACATGTTAGTGACAAGTTGTATATCGAGGTTGTAATGTTGTAT
TATACAAGAACATTGTAAAATATTTATAGTATATTGCAGCCAATATTTTT
TGAAAGGTGGGTGGATCGATGGCTTATGCTATCGATCAATCAGGAATTCA
ATTTAGAGTACTTAATAGTAGCAAAGGAGCTGCTGTTAGAGCAACACGTG
CTCAGGCAGATAAAATATTATATCGTCAAGCAATACGTAGTATTCTTGAA
TATCAAAAATTTTTGTTGGTTATTCAAGCGTCAGTAGAAGATTTAATAGT
TAGTGGGAACAAGATTGTTGGAGTAATTACTCCAAAATTAGGAATGAAAT
TTAGTGGTACGTCTGTTGTGTTGACAACCGGAACCTTTCTCAATGGTAAG
ATTCATATTGGGATGAATAATTTTAGAGGAGGTCGATCTGGAGATTCGGA
GTCATCGTCATTGTTATCAGAGCGATTGAAAGAATTGTCTTTTCAGATTA
GTCGGTTAAAAACGGGTACTTCTCCTCGTGTGCATACCAAAGGGATAAAT
TTTGGTTCTTTACGTGCACAATATAGTGATGATCCCATTCCTGTGTTTTC
ATTTATAGGATCTACAAAACTACATCCTACGCAGGTGCCT
DNA sequencing Why? Identifies Organisms How?
Dideoxy Sequencing Introduces Biological
Machines Where? Universities, MBL Societal
Impact? Forensics, Insurance
DNA detective