Serial dilutions (10−1, 10−3, 10−5) were prepared from these suspensions, which 100 μL of each one, were inoculated in different bacterial culture media, in duplicate.
We had an initial hit rate of 0.23%: 29 out of 12,800 compounds caused decreased growth in –Ade media without a significant growth decrease in +Ade media in duplicate well plates (Fig. 4).
3500 mouse SF cells transfected with various siRNA were seeded in 100 μl of media in duplicates in a 96X E-Plate, and proliferation was monitored in real time.
140 μL of the reconstituted βNAD+ was added to 6 μL of sample, lactate standards, sand pre-culture media controls in duplicate and incubated at 37°C for 30 minutes before the absorbance was recorded at 340 nm.
1.0×106 cells in 100 µl of media were transferred in duplicate wells to a 96-well plate.
Levels of IL-1β, TNF-α, and IL-6 in macrophage culture media were measured in duplicate in 3 separate assays using a bead-based immunoflourescence assay (Luminex Inc., Austin, TX, USA).
Cells were seeded into semi-solid media (1 ml, in duplicate) at the following densities: 4×104 bone marrow cells/ml and 5×105 spleen cells/ml for myeloid clonogenic progenitor assays, and 2×105 bone marrow cells/ml and 2×106 spleen cells/ml for B-lymphoid clonogenic progenitor assay.
Total NO, MMPs, TIMP-1, and C1-2C levels in the conditioned media were measured in duplicate.
Larvae were treated with 5 μM CM-H2DCFDA for 90 minutes and fluorescence of the media was measured in duplicate at 485 nm excitation/538 nm emission wavelengths on a SpectroMax M5e Multi-Mode Microplate Reader (Molecular Devices, Sunnyvale, CA).
Proteins that co-IPed with RNAs very different than the Mocks were grown in minimal media and re-purified in duplicate as described above.
Briefly, 100 μl or 200 μl of each sample (200 μl or 400 μl of the matched FBM-solid fraction to account for dilution with collagenase) was directly plated into a 10 cm diameter petri-dish (Corning Life Sciences, Amsterdam, Holland) with 15 ml of non-haematopoietic media (Miltenyi Biotec, Bisley, UK) in duplicate.
