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With five weeks' passage, the fateful encounter between a black youth who wanted to go to college and a Hispanic man who wanted to be a judge has polarized the nation.
Untransformed 16HBE cells (controls) and Cd-transformed cells at the 35th (5 μmol L−1 Cd for 14 weeks) passage were cultured in RPMI-1640 containing L-glutamine (Sigma, St Louis, USA), 10% fetal bovine serum (FBS), and 1% penicillin/streptomycin (Life, California Carlsbad, USA) at 37°C in a 5% CO2 humidified atmosphere.
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The stem cells were maintained for 3 4 weeks (passages 2 3) before proceeding to the surgical interventions.
The labeled cells were then allowed to grow for 4 6 weeks (passaging when confluent, and retaining all cells through the period), and were analyzed and sorted using a BD FACS Aria (VWR).
After a seasick six-week passage in a stripped-down troopship, the four women arrived back in England on Fay's 15th birthday -- and began a further round of camping out in rented flats, depending on hardscrabble attempts to earn a living.
"The commander turned out to be tremendous," said Mrs. Laychak, looking back on a harrowing 12-week passage through shock, panic and grief in which, the widow gratefully notes, Commander BrownWahler was always on call at all hours with a tonic mix of maternal compassion and executive resolve.
The other half was cultured again without treatment until confluence for 1 week (passage-1 chondrocytes).
To detect virus deletion variants with much greater sensitivity, Illumina NGS was performed on these five TEV lineages evolved for four 3-week passages.
Primary tumor cells were cultured for 1 week, passaged into a new 10-cm culture dish and allowed to grow to confluence.
Moreover, these effects can occur on relatively short timescales (i.e., deletion variants were generated de novo and reached frequencies of ∼ 0.25 within four 3-week passages).
This same mutation has also been previously reported in serial-passage evolution experiments of TEV in N. tabacum: in 10% of lineages after 15 one-week passages (Bedhomme et al. 2012), and in 30% of lineages after three 9-week passages (Zwart et al. 2014).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com