Hippocampal RNA Extraction and cDNA Synthesis

RNA isolation and reverse transcription were performed as described previously [65 (link)]. Briefly, total RNA was extracted from dorsal hippocampi using the Direct-zol^TM RNA MiniPrep (Zymo Research, Irvine, CA, USA) according to the manufacturer’s instructions [66 (link)]. cDNA was synthesized using the iScript kit (Bio-Rad Laboratories), which contains a blend of oligo(dT) and random hexamer primers in the iScript Reaction Mix, according to the manufacturer’s instructions. The specificity of PCR products was assessed by melting curve analysis.

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Ravanelli F., Musazzi L., Barbieri S.S., Rovati G., Popoli M., Barbon A, & Ieraci A. (2023). Differential Epigenetic Changes in the Dorsal Hippocampus of Male and Female SAMP8 Mice: A Preliminary Study. International Journal of Molecular Sciences, 24(17), 13084.

Publication 2023

Direct-zolTM RNA MiniPrep iScript kit

Cdna Hippocampi Isolation Oligo Primers Reverse transcription

Corresponding Organization : Università degli Studi eCampus

Other organizations : University of Milan, University of Milano-Bicocca, Centro Cardiologico Monzino, University of Brescia

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Variable analysis

independent variables

RNA isolation and reverse transcription

dependent variables

Specificity of PCR products

control variables

Total RNA extraction from dorsal hippocampi using the Direct-zol^TM RNA MiniPrep (Zymo Research, Irvine, CA, USA) according to the manufacturer's instructions
CDNA synthesis using the iScript kit (Bio-Rad Laboratories), which contains a blend of oligo(dT) and random hexamer primers in the iScript Reaction Mix, according to the manufacturer's instructions

controls

Melting curve analysis to assess the specificity of PCR products

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