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Various concentrations of recombinant human VEGF165 (200 and 500 ng) were added and incubated with HUVEC cells for 72 h.
Increasing amounts of each cDNA plasmid (3.3, 10, 30, 90 ng) were added to each well.
When higher levels of TBP (10 ng and 30 ng) were added, X gene transcription was further activated.
OCD3, 700 ng, were added to the reconstituted extracts (1 mL during these experiments) prior to injection on the LC-ESI-MS/MS.
Subsequently, fibrinogen (500 ng, Calbiochem) and uPA (10 ng) were added and incubated at 37°C for 0.5, 1, 2, 4 and 6 h.
DNA samples (200 ng) were added to the PCR reaction mixture (10 µl) containing 1 µl of 10× PCR buffer, 0.3 µl of 50 mM MgCl2, 0.2 µl of 10 mM dNTP each, 0.6 µl of DMSO, 0.14 µl of 5 U Platinum Taq enzyme (Invitogen corp ., 0.12 µl of 350 µg/ml primer mix (1∶1), and 7.64 µl of DNA in water.
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For each sample a 5.5 μL RLMM aliquot was dispensed into the wells and the 5.5 μL of the diluted sample genomic DNA (50 ng) was added.
DNase-treated RNA (100 ng) was added as template to SuperScript One-Step RT-PCR using Platinum Taq protocols to yield the corresponding cDNA.
One microliter of DNA (~25 ng) was added to 50 μl reaction volume containing 25 μl of Taq PCR mix (Generay, China), 23 μl dd water and 5 pmoles of each primer.
PA63 heptamer (15 ng) was added to the cis compartment.
Probe (0.5 ng) was added to each sample and incubated for 20 minutes on ice.
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