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In the "steps" section, for instance, users are walked through its nine phases of a successful flirtation, from the initial approach to asking for a person's phone number.
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For the more experienced, the Next Steps section looks at older records.
The steps followed for the numerical simulation are presented in "Simulation steps" section.
After these steps, sections were rinsed for 30 min in KPBS and incubated for 1 h in biotinylated goat anti-rabbit immunoglobulin G (IgG) or biotinylated horse anti-mouse IgG diluted 1 200 in KPBS containing 3% normal goat serum or normal horse serum.
Between steps, sections were washed twice for 10 min with TBS (0.05 M Tris buffer of pH 7.6 containing 0.3 M NaCl) and after step 4 with distilled water.
After repeated washing steps, sections were incubated with secondary antibodies for 1 h at RT.
Nodes were then submitted to microscopic evaluation: 5 10 microsections at representative levels of each sentinel lymph node were obtained by a step sectioning technique for nodes >5 mm.
Be well prepared for insults and remarks, read the Steps and Tips section for more info.
For reasons of practicality, five stepped sections were quantified for all markers within Studies A and B. Exploration of the data for all transformed markers revealed them to be log-normally distributed.
In the next step, sections were incubated for 48 h at 4°C with primary antibodies diluted in blocking solution.
Specimens from three-step sections were used for DNA extraction using DEXPAT (Takara, Tokyo, Japan) according to the manufacturer's instructions.
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