DNA extraction and fpg Assay：DNA的提取及空腹血糖测定.docVIP

Manuscript 2003-10-11408B Supplementary Methods Postmortem and Biopsy Cases Detailed case information is provided in Supplementary Table 1. The postmortem brain tissue samples used in this study were neuropathologically normal for age, and were derived from non-demented individuals. Some cases had been neuropsychologically tested as part of aging studies (77, 80, 82, 87, 88, 90(B) and 91 years old; Supplementary Table 1). Tissue was procured in accordance with institutional guidelines. Human frontal cortical grey matter samples were dissected from the frontal pole (Brodmann area 10), and were snap frozen in liquid nitrogen and stored at -85oC. Some intracortical biopsy samples were also included in this study. Cluster and correlation coefficient analysis (Fig. 1a, b) utilized 30 cases (Supplementary Table 1; 26, 26B, 27, 29, 30, 36, 37, 38, 40, 42, 45, 48, 52, 53, 56, 61, 66, 70, 71, 73, 77, 80, 81, 85, 87, 90, 90B, 91, 95 and 106 years old). Group comparison (Table 1 and Supplementary Table 2) utilized cases ≤42 years old (26, 26B, 27, 29, 30, 36, 37, 38, 40 and 42 years) and ≥73 years old (73, 77, 80, 81, 85, 87, 90, 90B, 91, 95, and 106 years). RNA Isolation and Microarray Hybridization Dissected cortical grey matter was cut into small pieces in the frozen state and ~70 mg was homogenized immediately in Trizol (Gibco) and RNA was isolated. RNA that was intact by electrophoresis and had an A260/A280 ratio ≥ 1.9 was used for cDNA synthesis. cDNA, cRNA synthesis, cRNA fragmentization and preparation of the hybridization cocktail were carried out according to the Affymetrix protocol. After hybridization for 16 hrs at 45?C in the Genechip hybridization oven 640 (60vrpm), the probe arrays were washed,