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He escaped but was quickly picked up again, and transferred to the notorious slave labour camp of Dora.
After the final wash, the beads were pelleted again and transferred to a new tube for elution.
After 72 hours, EB were harvested again and transferred to « hematopoietic medium » consisting of StemPro media supplemented with 100 ng/ml human recombinant VEGF (R&D Systems), 5 ng/ml bFGF, 100 ng/ml SCF (kind gift from Amgen), 100 ng/ml FLT3-L (kind gift from Amgen) and 40 ng/ml TPO (Peprotech, UK) for 4 additional days.
All grids were rinsed in PBS, fixed using 1% gluteraldehyde for five minutes, rinsed again and transferred to a UA/methylcellulose drop before being collected and dried.
Samples were then washed again and transferred to cluster tubes in 4% paraformaldehyde in PBS (400 μl) and acquired using an LSRII flow cytometer (BD Biosciences) set up with the beads provided and the highest concentration standard; the stopping gate was set so that at least 300 events were collected for each sample.
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Stir again and transfer to serving dish.
Add eggs and sugar, pulse briefly, add 1 1/2 cups milk, pulse again and transfer to heavy saucepan.
After a short period in charge he was again promoted and transferred, taking up the role of Staff Captain on the liner RMS Empress of Australia.
The coverslips were again washed and transferred to an object slide with a drop of ProLong Gold antifade reagent for mounting.
After o/n incubation in an orbital shaker at 300 rpm, one ml of the culture was again washed and transferred into 10 ml MMES as described above.
They are then disinfected with iodine and/or formalin, rinsed again, counted and transferred to holding tanks or directly to larval rearing tanks.
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