Telomeric DNA damage response activation mediates Abeta42 oligomers neurotoxicity in Alzheimer's disease
Telomeric DNA damage response activation mediates Abeta42 oligomers neurotoxicity in Alzheimer's disease
Mouse primary neurons were processed after 48 hours for immunofluorescence, RNA extraction, and viability assay﻿. Maxwell® RSC simplyRNA Tissue Kit (AS134, Promega) For each sample, 500 ng of total RNA were used to generate a library of fragments using Illumina Stranded mRNA prep ligation kit. Oligo(dT) magnetic beads purify and capture the mRNA molecules containing polyA tails. The purified mRNA was fragmented and copied into first strand complimentary DNA (cDNA) using reverse transcriptase and random primers. In a second strand cDNA synthesis step, dUTP replaces dTTP to achieve strand specificity. The final steps added adenine (A) and thymine (T) bases to fragment ends and ligate adapters. The resulting products were purified and selectively amplified with 12 cycles of PCR to generate an indexed library of fragments. The library was quantified using Qubit 4.0 fluorimeter, checked on Agilent Bioanalyzer 2100