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Standard I Medium was purchased from Merck (Zug, Switzerland).
The strain was grown on Standard I Medium as described previously (Corvini et al. 2004c).
Given and for a given shot, we classify each into three classes, namely, low (I), medium (II), and high (III), based on an equi probability classification.
Microorganisms were cultivated on a mineral medium (MM; Brunner, DSMZ no. 462) and, for purity control, on non-selective Standard I medium (Merck 7881, DSMZ no. 453) or nutrient broth (NB).
Cells were washed by replacing the complete medium by the Optimem I medium (50µl).
Lipofectamine - siRNA complexes were produced in Opti-MEM I medium (Invitrogen) according to the manufacturer protocol.
N2A cells were detached from growth plates using trypsin and then resuspended in Opti-MEM I medium (Invitrogen) at a concentration of 50 cells/µl.
For the next 2 days, cultures were fed with Epidermalization II medium, which is identical to Epidermalization I medium except that it contains 0.1% unchelated newborn calf serum.
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For each experiment, approximately 1 mg plasmids were pre-mixed with 3 mg PEIs in 50 mL fresh SMM 293T-I medium (Sino Biological Inc).
Lipofectamine Reagent and Optimem-I medium were purchased from Invitrogen.
Transfections were performed with 20% confluent HeLa cells using Oligofectamine (Invitrogen) in Optimem-I medium (Gibco) with 0.2 μM siRNA.
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