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Chunk #7 — Methods

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The relationship of 5HTT (SLC6A4) methylation and genotype on mRNA expression and liability to major depression and alcohol dependence in subjects from the Iowa Adoption Studies.
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DNA and RNA used in the studies were both prepared from the lymphoblast cell lines. These lymphoblast cell lines were prepared by standard EBV transformation [Klaus, 1987] and the media changed 24 hr prior to the use of the cell lines for the preparation of biomaterial. Total RNA was prepared from the lymphoblast using an Invitrogen RNA purification kit (Invitrogen, Carlsbad, CA) according to the instructions of the manufacturer [Bradley et al., 2005]. DNA was prepared for the cell lines using cold protein precipitation [Lahiri and Nurnberger, 1991]. Not all parameters were successfully measured in all cell lines. Hence, the “n” for each experiment may vary.