3. Preculture of Sf-9 cells.
4. Insect cell culture medium (e.g., Sf900 II, Thermo Fisher
Scientific, Waltham, MA, USA).
5. Temperature probe (e.g., Labfors 5 cell, Infors HT, Basel,
Switzerland).
6. pH and DO probe (e.g., EasyFerm/VisiFerm, Hamilton,
Bonaduz, Switzerland).
7. Glass bottles with connector caps and bottom drain for acid,
base, medium and inoculation.
8. Male and female Luer lock adapters.
9. Luer lock lids.
10. Sterile, hydrophobic air filter with Luer lock adapter (e.g.,
Minisart, Sartorius, Go ¨ttingen, Germany).
11. Sterile, single-packed, 10-mL syringe with Luer lock adapter
(e.g., B. Braun, Melsungen, Germany).
12. 1 M phosphoric acid.
13. 1 M sodium hydroxide.
14. 70% (v/v) ethanol.
2.4.6 Materials for Online
Dielectric Spectroscopy
1. Dielectric spectroscopy probe (e.g., InCyte, Hamilton, Bonaduz, Switzerland).
2. Signal transformation box (e.g., Cell Density ComBox, Hamilton, Bonaduz, Switzerland).
3. Documentation software (e.g., Cell Density software, Hamilton, Bonaduz, Switzerland).
4. Cleaning solution (15 g/L Na 2 SO 3 ).
2.5 Production of a
P1 Baculovirus stock
and Amplification
The required materials for the production of a P1 virus stock as well
as for the amplification of the virus stock are described in this part
of the chapter.
2.5.1 Materials
for the Transfection of Sf-9
Cells for the Generation
of a P1 Baculovirus Stock
1. Material for cell counting (see Subheading 2.4.2).
2. Spodoptera frugiperda cell suspension (2.5 mL) with a cell
density of 0.3 Â 10
6 cells/mL (e.g., TriEx Sf-9, Merck, Darmstadt, Germany).
3. Insect cell culture medium (e.g., Sf900 II, Thermo Fisher
Scientific, Waltham, MA, USA).
4. Isolated Bacmid DNA (e.g., Bacmid DNA of AcMNPV-Orf2PCV2; see Subheading 3.3).
5. Transfection reagent (e.g., TransIT
® -Insect Transfection
Reagent, Mirus Bio LL, Madison, Wisconsin, USA).
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