DNA Joining

Linkers and Adapters

C G A T G G A T C C A T C C
| | | | | | | | | | | | | |
G C T A C C T A G G T A G G
CH
2
Base
O
CH
2
Base
O
CH
2
Base
O
O
O
O
O
O
O
H
3
O
O
O
O
5
The structure of a polynucleotide strand
showing the chemical distinction
between 5-P and 3-OH termini.
C H
2
B a s e
O
C H
2
B a s e
O
C H
2
B a s e
O
O
O
O
O
O
O
H
3
O
O
O
O
5
5
3
3
5
In the double helix the
polynucleotide strands are
antiparallel
5
5 3
3 5
5
3
3
Ligation takes place between 5 P and 3 OH termini
5
5
PO
2-
3
PO
2-
3
OH
OH
HO
The modified 5 OH
terminus
Alkaline Phosphatase
HO
O H
OH
HO
DNA Ligase
Polynucleotide kinase
PO
2-
3
PO
2-
3
5 P terminus 5 P terminus
Ligation using Adapters
Modification in 5 terminus
with Alkaline Phosphatase
DNA Bases
Terminal transferase
dCTP
Synthesis of a homopolymer tail
Ligation of homopolymer tail
dGTP
The repair step
Nick
Discontinuity
DNA Ligase