3. For groups with electroporation, the mice need to be anesthetized by intraperitoneal injection of ketamine (75 mg/kg)
mixed with medetomidine (1 mg/kg).
4. After anesthesia, the electrode needles are inserted into the
tibialis posterior muscle at a depth of 3 mm and the impedance
between electrodes is measured to make sure the electrodes are
in the muscle. The electrodes are well positioned when the
measured impedance is between 1 and 2 kΩ.
5. The syringe needle is then inserted at the same depth, between
the two electrodes, and the injection of DNA vaccine is
performed.
Fig. 4 Example of agarose gel electrophoresis of DENV2 DNA vaccine Giga prep
Table 1
Standard mouse experimental design to assess different DNA vaccine formulations and effect of
adjuvants
Group number
No. of mice
Route
DNA vaccination
Protein boost
1
6
i.m.
a
2Â plasmid 1
2Â protein + adjuvant
2
6
i.m.
a
2Â plasmid 1 + adjuvant
2Â protein + adjuvant
3
6
i.m.
a + EP
b
2Â plasmid 1
2Â protein + adjuvant
4
6
i.m.
a + EP
b
2Â plasmid 1 + adjuvant
2Â protein + adjuvant
5
6
i.m.
a
–
4 Â protein + adjuvant
6
6
i.m.
a + EP
b
4Â plasmid 1 + adjuvant
–
7
6
i.m.
a + EP
b
2Â saline + adjuvant
2Â saline + adjuvant
a
i.m. injection of DNA at 2 μg/μL
b
Parameters: V ¼ 100, time ¼ 50 ms, interval ¼ 100 ms, number of pulses ¼ 3
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Lei Li et al.