GSM7440157: Old, female, biological replicate 2, technical replicate 2, ST-RNAseq; Mus musculus; OTHER
Tissue sections were first stained with Hematoxylin-Eosin (H&E), followed by cDNA synthesis, amplification, barcoding, and library construction (see Methods). Following this demonstrated protocol, we obtained high-quality spatially resolved RNA-seq data from these 24 samples and sequenced them on the NovaSeq™ 6000 platform achieving an average coverage of 171 million read pairs per sample 10 µl (25%) of the total amplified cDNA was used as input to perform enzymatic fragmentation, end repair and A-tailing (10X Genomics, PN1000196). A double-sided size selection was then performed with SPRIselect (Beckman, Coulter B23318). TruSeq read2 primer was added via adaptor ligation (10X Genomics, PN1000196). P5, P7, i7, and i5 sample indices were added via index PCR with the individual dual index TT set A (10X Genomics, 3000431). The cycle number for index PCR was calculated based on the yield of the 25% carry forward cDNA material and 15 cycles were used. A double-sided size selection of the post-index PCR sample was performed with SPRIselect (Beckman, Coulter B23318).