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Purified lysates, normalized for protein content using a BCA assay, were run on 12% Novex Tris-Glycine gels (Invitrogen) and transferred to nitrocellulose membranes (Whatman, Florham Park, NJ, USA).
The PCR reactions for each assay were run in triplicate and the results were averaged.
The Luminometric Methylation Assay (LUMA) and LINE-1 methylation assay were run on all samples in duplicate.
The reactions were incubated in a 96-well optical plate at 50°C for 2 min, at 95°C for 10 min, following by 40 cycles of 95°C for 15 seconds and 60°C for 1 min. The real-time PCRs for each assay were run in triplicate.
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The assay was run in triplicate.
The assay was run in triplicate for each sample.
Following exposure the MTT assay was run as previously described.
Each TaqMan assay was run in four replicates for each RNA sample.
A conventional genotyping assay was run in parallel, based on nested PCR amplification (Figure 6C).
Each assay was run in duplicate.
Assay was run on Mx3000P (Stratagene).
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