RNA Extraction and qPCR Analysis from Cells and Brains

Cells were lysed using the TRIzol reagent (Invitrogen). Brains were removed from euthanized mice and frozen in liquid nitrogen. RNA was extracted using the TRIzol reagent (Invitrogen) according to manufacturer. After DNaseI treatment (Invitrogen) RNA from cells and brains was reverse transcribed into cDNA with oligo d(T) primer and OMNISCRIPT RT KIT (Qiagen). QPCRs were performed as described (Piersanti et al., 2015 (link)) using the following primers:

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Burla R., La Torre M., Zanetti G., Bastianelli A., Merigliano C., Del Giudice S., Vercelli A., Di Cunto F., Boido M., Vernì F, & Saggio I. (2018). p53-Sensitive Epileptic Behavior and Inflammation in Ft1 Hypomorphic Mice. Frontiers in Genetics, 9, 581.

Publication 2018

TRIzol reagent DNaseI treatment OMNISCRIPT RT KIT

Brains Cdna Cells Frozen Mice Nitrogen Oligo Primers Trizol

Corresponding Organization : Nanyang Technological University

Other organizations : University of Turin

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Variable analysis

independent variables

TRIzol reagent (Invitrogen) for cell lysis
DNaseI treatment (Invitrogen)

dependent variables

RNA extraction
CDNA reverse transcription

control variables

Brains removed from euthanized mice
Brains frozen in liquid nitrogen

controls

Positive control: Not specified
Negative control: Not specified

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