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"As a courtesy for the inconvenience, please accept a voucher for three additional hours of your life, redeemable upon your death".
The low-light treatment (low-light relative to the maximum light intensity) consisted of a 16-h day as in the rearing cage (described above) followed by a sudden increase to ~300 lux for three additional hours at sunset (Fig. 1).
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The temperature was reduced to 16°C for 1 h at which time recombinant SpGH38 was induced by the addition of 0.2 mM isopropyl 1-β-D-thiogalactopyranoside and incubated for sixteen additional hours.
In the most comprehensive account to date of the missing Malaysia Airlines Flight 370's fate, Malaysian Prime Minister Najib Razak on Saturday said that the plane was steered off course after its communications systems were intentionally disabled and that it could have flown for seven additional hours.
The obtained suspension was magnetically stirred for two additional hours at 80 °C.
The culture was diluted 1 10 in fresh 3.7% BHI and incubated for two additional hours to reach log-phase growth.
As part of the agency's contract with its operators, if a driver works on his or her birthday, he or she is automatically credited for eight additional hours.
50 µl of Dynabeads® M-270 Streptavidin (Dynal Biotech, Invitrogen) pre-washed twice and pre-equilibrated for 2 h at 4°C with blocking buffer (base buffer with 100 mM NaCl, 100 units/ml RNasin, 0.2 mg/ml tRNA, 0.1 mg/ml glycogen, 10 mM BSA and 2 mM DTT) was added to the binding mixture and incubated for two additional hours.
Expression of target proteins was induced by nisin (see Results part for dilution used; for every round of nisin production, systematic tests were performed to determine optimal dilutions required for production of target proteins) and cultures were incubated for four additional hours.
For cytotoxicity assays, C-1330 concentranging ranging from 10 nM to 100 μM were added for two additional hours.
Cells were grown at 37°C until OD600 nm 0.5 then induced with 0.5 mM IPTG for five additional hours.
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