Page 34 - Genetics_From_Genes_to_Genomes_6th_FULL_Part3

FEATURE FIGURE 9.7

Automated Sanger Sequencing

(a) Hybridization of template and primer. A cloned recombinant these strands serves as the template. The recombinant DNA
DNA is denatured (melted into single strands), using heat to is mixed with an oligonucleotide primer (previously made in a
break apart the hydrogen bonds joining the two strands. One of DNA synthesizer) whose sequence is complementary to about

(a)
Vector

Recombinant plasmid

Human
DNA insert

Melt into single strands
Hybridize with primer

Template
Insert
Vector

G C T C A G
A G C G A G T C A C C G T C G A G T C G 5'
3'
+ DNA polymerase
Primer
5' + dATP, dCTP, dGTP, dTTP
3' + ddATP, ddCTP, ddGTP, ddTTP
(b) – Red dye
H
G C T C A G T Dideoxynucleotide Gel 3'
5' H 3'
G
– Yellow dye A
H C
G C T C A G T G T
5' H 3' 3'
C
H G
G C T C A G T G G
5' H 3' 3' A
– Purple dye C Direction of electrophoresis
H
G C T C A G T G G C
5' H 3' 3' G
– Green dye
H G
G C T C A G T G G C A
5' H 3'
T
H
G C T C A G T G G C A G Detector
5' H 3'
5'
Primer Complementary to (Sequence of
template strand of newly synthesized
insert
DNA)

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