GSM9473146: 7 Month, 5xFAD negative, rep 2; Mus musculus; RNA-Seq
To isolate the CA2 area of the hippocampus for the bulk sequencing, we prepared 500 µm acute coronal brain slices as described previously.25 Slices were transferred to a Petri dish containing cold carbogen-bubbled (95% O2 + 5% CO2) cutting solution. Slices with hippocampus were visualized under a dissection microscope, and the CA2 area was anatomically identified by the neurons with larger cell bodies than CA1 neurons and with an intermediate cell density between loosely packed CA3 and more densely packed CA1 neurons.26 Using a tissue puncher (Ted Pella #15116-1 ID 1 mm), we isolated the CA2 area and snap-froze it. All samples were immediately lysed and homogenized using plastic tissue grinders before being stored in pre-chilled tubes at −80°C for a minimum of 24 h. RNA isolation was carried out on thawed samples using a TRIzol Plus RNA Purification Kit (Cat#12183555, Invitrogen). Following purification and nanodrop sample verification, samples were sent to the Yale Center for Genome Analysis (New Haven, CT), where they underwent mRNA (Poly A) library preparation and sequencing on an Illumina NovaSeq 6000.
GEO
GSM9473146_r1