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Bisulfite treatment was carried out on genomic DNA using the EZ DNA Methylation-Gold Kit® (Zymo Research, Orange, CA) according to the manufacturer's protocol with minor modifications detailed below [17].
ChIP was performed as previously described (Lim et al., 2009) with minor modifications detailed below.
Amplex Red assay kit was purchased from Invitrogen (cat. no. A12219) and used with modifications detailed below.
Real time quantitative PCR (RT-qPCR) was performed as described in Iskandar et al. [ 16] with the modifications detailed below.
This involved obtaining approval for intervention units to have physical work environment modifications (detailed below) as well as other intervention activities at the organisational and individual levels.
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Detailed below are additional modifications that the user may select to further enhance the gene design and arrive at a highly optimized sequence.
Similarly, we failed to confirm other positive reports of associations with hormonal risk factors, or of effect modification, as detailed below.
The specific modifications and the items changed are detailed below.
Although the work is definitely of eLife quality, the manuscript would benefit from some modifications, further experiments and reorganization, as detailed below: Major comments: 1) We are not sure if the microarray/ChIP-Seq analysis adds much to the paper.
The electron proton distances were calculated from the intensity ratios (Iox/ Ired) as described previously, including the modifications introduced by Allison et al. as detailed below.
This was performed as described (Teismann et al., 2003) with the minor modification that incubation with primary antibodies was performed sequentially as detailed below.
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