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After irradiation at 60 Gray in a Torrex 150D (Astrophysics Research Corp., City of Industry, CA) embryos from both collections were aged for 20 min on the apple-juice agar plates and mixed before fixation.
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Phase I, II, and III collections were aged 1.5, 2.5, and 3.5 hours respectively following the one hour collection and then stopped.
Embryos from 0 2 hr egg collections were aged for 1 hr at 25°C and were fixed and stained as previously described (Stevens et al., 2009).
We recruited all the students of other Forms (n = 2,251) who were present on the day of the data collection, were aged 15 24 years, and agreed to participate in the study.
For PS6, a 6 hr collection was aged for a further 4 hr.
For this, we analyzed staged embryonic collections from 0-2 hr that were aged for 3 hr before fixation.
In the following data collection phases (when the children were aged 6 9 months, 12 16 months, and 2 3 years), the mothers were re-contacted and invited to participate again.
In order to obtain a time series of embryos, the eggs from the 2-h collections were then aged for the required time (between 4 22 h) at 25°C.
Control animals did not exhibit any BSE symptoms at the time of collection and were age matched with BSE cases from the same farm.
Serum collections like these would in general be age-stratified, but we do not take advantage of the age information in the analysis presented here.
Additionally, these desiccation methods are commonly used in the field, and existing collections (dried using these methods) could be age-graded by NIRS.
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