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Scientific Reading and Writing in English Shu-ying Wang, Ph.D. Dept. Microbiology and Immunology Tel: +886-6-2353535 ext. 5634 Fax: +886-6-2082705 Email : [email protected] Scientific Reading and Writing in English Fall, 2009 Time: 1:10-3:00 pm, every Tuesday
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Time: 1:10-3:00 pm, every Tuesday
Place: Conference room (82-1124), Dept. Microbiology and Immunology (11th floor)
is to address
“what/how you did it”.
“MMs”. The “MMs” section should include sufficient technical information to allow the experiments to be repeated. When centrifugation conditions are critical, give enough information to enable another investigator to repeat the procedure: make of centrifuge, model of rotor, temperature, time at maximum speed, and centrifugal force ( X g rather than revolutions per minute). For commonly used materials and methods (e.g., media and protein concentration determinations), a simple reference is sufficient.
in a table footnote or figure legend.
Cells, viruses, and reagents. The B-cell line Raji was maintained in RPMI medium containing 10% fetal bovine serum according to American Type Culture Collection instructions. DV2 strains PL046 and M16681 were propagated in C6/36 mosquito cells and titrated on BHK cells as previously described (18). Human DV3 immune serum was obtained with consent from an infected patient. The titer of this serum was 1:12,000 against DV3 and 1:3,200 against DV2 strain PL046, as determined by measuring 50% plaque reduction in a neutralization assay using BHK cell monolayers (14). Control serum was collected from a healthy blood donor without DV-specific antibodies in serum as determined by an enzyme-linked immunosorbent assay (ELISA) as previously described (19).
Flow cytometry. Infected B cells were washed with phosphate-buffered saline (PBS) twice, fixed, permeabilized, washed, and incubated with the IgG fraction of NS1 hyperimmune serum or normal mouse serum. Cells were then washed and incubated with fluorescein isothiocyanate-conjugated goat anti-mouse IgG (Endogen). After incubation, cells were washed twice with PBS, resuspended in PBS, and then analyzed with a FACScan flow cytometer (Becton Dickinson Immunocytometry Systems, San Jose, Calif.) as previously described.
However, related methods should be described together.