Please find the following protocol:

Baculovirus Transfection Using Calcium Chloride

1. Seed 2 million Sf9 cells into a 25 cm2 T-flask and allow cells
to attach for at least 1 hr.

2. Mix 1 μg of the insect baculovirus vector AcMNPV linear viral
DNA and 2 μg of recombinant plasmid DNA in a snap cap
polypropylene tube.

3. Prepare 5 ml of HEBS/CT by combining 0.5 ml of 10X HEBS,
75 μl of sonicated calf thymus DNA, 4.425 ml of ddH2O and
adjust the pH to 7.05 to 7.1 with 0.1 M NaOH. Filter sterilize
and allow the solution to come to room temperature before
use.

4. Add 950 μl of HEBS/CT to the DNA mixture and mix by
vortexing.

5. Add 50 μl of 2.5 M CaCl2 to the DNA/HEBS/CT mixture
drop-wise with constant mixing.

6. Incubate at room temperature for 30 min.

7. Remove the media from the tissue culture T-flask and replace
it with 2 ml of Complete Grace's Insect Medium.

8. Add DNA precipitate solution to the cell culture flask
drop-wise with mixing and incubate at 37°C for 4 hr.

9. Remove the Grace's media from the flask. and rinse the cells
once with Complete Grace's Insect Media.

10. Remove the Grace's Insect Media and replace it with
Complete TNM-FH Insect Media.

11. Incubate at 27°C for 3 to 6 days, looking for signs of
infection daily. Some protocols state that the infection
takes only 2 days, but with this protocol it actually takes
longer.

12. Harvest the supernatant when the cells are ready and store
the transfection supernatant at 4°C.


Solutions

TNM-FH Insect Medium (Complete).........TNM-HM Insect Medium
.................100 U/ml Penicillin
....................2 mM Glutamine
............10% Fetal Calf Serum
.........100 μg/ml Streptomycin
Grace's Insect Medium (Complete)...................2 mM Glutamine
............10% Fetal Calf Serum
...........Grace's Insect Medium
............100 Units/ml Penicillin
.........100 μg/ml Streptomycin
0.1 M NaOH
CT DNA............................1 mg/ml Sonicated Calf Thymus DNA
2.5 M CaCl2.................Make 0.5 ml aliquots and store at -20°C
10X HEBS Buffer...............................200 mM HEPES, Free Acid
.....7 mM Sodium Phosphate Dibasic (Na2HPO4)
.............................................600 mM Glucose
..........Adjust the pH to 7.1 with fresh 5 M NaOH
........................Sterile filter and store at -20°C
...................................................1.37 M NaCl
.....................................................50 mM KCl


BioReagents and Chemicals

Fetal Calf Serum
AcMNPV viral DNA
Sodium Hydroxide
Glucose
HEPES, Free Acid
TNM-FH Insect Medium
Grace's Insect Medium
Sodium Phosphate, Dibasic
Potassium Chloride
Glutamine
Sodium Chloride
DNA, Calf Thymus
Streptomycin
Penicillin
Calcium Chloride


Reference Link:

http://www.bio.com/protocolstools/protocol.jhtml?id=p605