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The enzyme reactions were continued for one hour on an orbital shaker at 37°C before being stopped by addition of 1.25 ml 200 mM glycine solution at pH 10.4.
Resuscitation was performed by infusing 2x shed blood volume in normal saline over 30 min, followed by 1/2 shed blood volume returned over 30 min, then completed with 2x shed blood volume in normal saline over 60 min. Lymph collection continued for one hour post completion of resuscitation and all lymph samples were then centrifuged at 5000 × g for 10 min to remove cellular components.
Tissue homogenates were added 24 hours later and incubation was continued for one hour.
In case of neutral pH controls cultivation was continued for one hour in standard BHI broth.
Continued for one hour per day, heat shocks of otherwise null mothers produced significantly less ovarian BicD than did wild-type mothers (Fig. 1C).
60 µl of N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) and 6.0 µl of chlorotrimethylsilane (TMS-Cl) were added and the reaction was continued for one hour.
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Incubation was continued for one additional hour for one of the infected/control class of cells, a total of 2 h incubation, and for 5 more hours for the other class, a total of 6 h incubation.
Reduce temperature to 300 degrees and continue baking for one hour and 10 minutes, until top is puffed and golden -- some cracking is to be expected -- and a knife inserted in center comes out clean.
Medication was continued for one month.
The therapy was continued for one year.
TQ treatment was continued for one month.
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