4. Inoculate the small scale culture 1:500 into 650 mL 2YT
medium in a 2 L shaker flask to make the total culture volume
of 2.5 L for a Giga prep. Shake overnight as above.
5. Pellet the bacteria and perform Giga prep as per the manual of
Qiagen Endotoxin Free Plasmid Purification kit. The expected
yield is generally 7–10 mg (see Note 4).
6. Measure plasmid DNA concentration using NanoDrop or
fluorescence-based method and also perform agarose gel electrophoresis to confirm the supercoiled plasmids (Fig. 4).
3.2 Mouse
Vaccination
1. Prepare vaccine formulation according to Table 1 in sterile
tubes. Select any desired adjuvant to assess its effects on DNA
vaccine immunogenicity.
2. Inject 50 μL of vaccine intramuscularly for all mice that do not
need electroporation.
Fig. 3 Plasmid map of DENV2 DNA vaccine
DNA Vaccine Design and Adjuvant Selection
413
medium in a 2 L shaker flask to make the total culture volume
of 2.5 L for a Giga prep. Shake overnight as above.
5. Pellet the bacteria and perform Giga prep as per the manual of
Qiagen Endotoxin Free Plasmid Purification kit. The expected
yield is generally 7–10 mg (see Note 4).
6. Measure plasmid DNA concentration using NanoDrop or
fluorescence-based method and also perform agarose gel electrophoresis to confirm the supercoiled plasmids (Fig. 4).
3.2 Mouse
Vaccination
1. Prepare vaccine formulation according to Table 1 in sterile
tubes. Select any desired adjuvant to assess its effects on DNA
vaccine immunogenicity.
2. Inject 50 μL of vaccine intramuscularly for all mice that do not
need electroporation.
Fig. 3 Plasmid map of DENV2 DNA vaccine
DNA Vaccine Design and Adjuvant Selection
413