Research plan for INTEL ISEF
Posted: Sun Dec 09, 2012 4:47 pm
Hi,
I am presently writing down my research plan to apply for INTEL. I have a question about it. In the instructions, it is written that the procedures should be written in DETAILS. I am not sure if what I have written is detailed enough, because I don't want it to be too long either. For example, here a paragraph:
Quantification of IFN-γ by ELISA
Levels of IFN-γ secreted by C57BL/6 cells were measured using an ELISA sandwich assay. Briefly, 96-well ELISA plates were coated with 100 µl/well of anti-IFN-γ antibody diluted (1/250) in 0.1M sodium carbonate buffer, pH 9.5 for 16 hr at 4°C. The plate was washed with phosphate buffered saline (PBS) containing 0.05% Tween (0.05% Tween/PBS), and the plate was blocked with PBS containing 10% FCS (10% FCS/PBS) (1h, 25°C). The plate was washed again with 0.05% Tween/PBS, and 100 µl of supernatant from C57BL/6 cells or IFN-γ standard, diluted in 10% FCS/PBS in duplicate were added and incubated (2h, 25°C). The plate was washed again with 0.05% Tween/PBS, and a dilution of biotinylated anti-mouse IgG antibody and HRP-conjugated streptavidin were added to each well (1h, 25°C). After washing, TMB substrate solution (100 µl/well) were added, and the reaction was stopped after 30 min. Absorbance was measured at 450 nm using an ELISA reader (Bio-Tek Instruments Inc., Winooski, VT).
In this paragraph, I haven't explained how the standards are prepared. Is it important or is it detailed enough?
Thank you!
Minuoja
I am presently writing down my research plan to apply for INTEL. I have a question about it. In the instructions, it is written that the procedures should be written in DETAILS. I am not sure if what I have written is detailed enough, because I don't want it to be too long either. For example, here a paragraph:
Quantification of IFN-γ by ELISA
Levels of IFN-γ secreted by C57BL/6 cells were measured using an ELISA sandwich assay. Briefly, 96-well ELISA plates were coated with 100 µl/well of anti-IFN-γ antibody diluted (1/250) in 0.1M sodium carbonate buffer, pH 9.5 for 16 hr at 4°C. The plate was washed with phosphate buffered saline (PBS) containing 0.05% Tween (0.05% Tween/PBS), and the plate was blocked with PBS containing 10% FCS (10% FCS/PBS) (1h, 25°C). The plate was washed again with 0.05% Tween/PBS, and 100 µl of supernatant from C57BL/6 cells or IFN-γ standard, diluted in 10% FCS/PBS in duplicate were added and incubated (2h, 25°C). The plate was washed again with 0.05% Tween/PBS, and a dilution of biotinylated anti-mouse IgG antibody and HRP-conjugated streptavidin were added to each well (1h, 25°C). After washing, TMB substrate solution (100 µl/well) were added, and the reaction was stopped after 30 min. Absorbance was measured at 450 nm using an ELISA reader (Bio-Tek Instruments Inc., Winooski, VT).
In this paragraph, I haven't explained how the standards are prepared. Is it important or is it detailed enough?
Thank you!
Minuoja