Nucleic Acid Based Nanoconstructs for Environmental Analysis …
583
(continued)
Name
Oligonucleotide sequence
Modification
S2-20
GTCGCAGAGTTGACCTGACC
S3-20
ACTCTTCTCGCCACCTCACC
S1
GGTGAGGTGGCGAGAGCGACGATCCATGCC
S2
GGCATGGATCGTCGCAGAGTTGACCTGACC
S3
GGTCAGGTCAACTCTTCTCGCCACCTCACC
ss1
CAGATCCGAGGAATCTG
ss1c
CAGATTCCTCGGATCTG
ss2
ACATGAAATTAAATACATGT
ss2c
ACATGTATTTAATTTCATGT
Nanobio-Construct Design
Our nanobio-construct comprises a three-way junction with biotinylation at different
locations to enable attachment with avidin. The constituent components include the
following:
A. Three-Way Junction (3WJ) comprising constituents as shown below
B. Biotinylated Three-Way Junctions
• 3WJ1 one arm bearing shorter sequence modified with biotin.
• 3WJ2 two arms bearing shorter sequences modified with biotin groups.
The three-way junctions (3WJs) were prepared by incubating the appropriate
DNA strands in a 10 mM Tris-Cl (pH 7.5) buffer containing, 1 mM EDTA and 1 M
NaCl. The concentration of DNA was 2 μM of each strand. The mixture was heated
for a brief period (at ~95 °C) and slowly cooled to room temperature. Based on the
above architectures, two types of nanoconstructs were constructed. These were borne
from 3WJ1 and 3WJ2 by the addition of avidin.
M3WJ1: Matrix 3WJ1 (M3WJ1) was prepared by incubating of avidin with biotinylated 3WJ1 in a 1:3 stoichiometry at room temperature for 30 min in a 10 mM Tris-Cl
(pH 7.5) buffer containing, 1 mM EDTA, and 1 M NaCl.
583
(continued)
Name
Oligonucleotide sequence
Modification
S2-20
GTCGCAGAGTTGACCTGACC
S3-20
ACTCTTCTCGCCACCTCACC
S1
GGTGAGGTGGCGAGAGCGACGATCCATGCC
S2
GGCATGGATCGTCGCAGAGTTGACCTGACC
S3
GGTCAGGTCAACTCTTCTCGCCACCTCACC
ss1
CAGATCCGAGGAATCTG
ss1c
CAGATTCCTCGGATCTG
ss2
ACATGAAATTAAATACATGT
ss2c
ACATGTATTTAATTTCATGT
Nanobio-Construct Design
Our nanobio-construct comprises a three-way junction with biotinylation at different
locations to enable attachment with avidin. The constituent components include the
following:
A. Three-Way Junction (3WJ) comprising constituents as shown below
B. Biotinylated Three-Way Junctions
• 3WJ1 one arm bearing shorter sequence modified with biotin.
• 3WJ2 two arms bearing shorter sequences modified with biotin groups.
The three-way junctions (3WJs) were prepared by incubating the appropriate
DNA strands in a 10 mM Tris-Cl (pH 7.5) buffer containing, 1 mM EDTA and 1 M
NaCl. The concentration of DNA was 2 μM of each strand. The mixture was heated
for a brief period (at ~95 °C) and slowly cooled to room temperature. Based on the
above architectures, two types of nanoconstructs were constructed. These were borne
from 3WJ1 and 3WJ2 by the addition of avidin.
M3WJ1: Matrix 3WJ1 (M3WJ1) was prepared by incubating of avidin with biotinylated 3WJ1 in a 1:3 stoichiometry at room temperature for 30 min in a 10 mM Tris-Cl
(pH 7.5) buffer containing, 1 mM EDTA, and 1 M NaCl.