title: GSM8754374: pic43_Aged_replicate 1-4; Nothobranchius furzeri; OTHER
library_construction_protocol: Lysis solution(0.1% Tween and 400 ug/ml of ProteinaseK in PBS) were loaded onto the specimens and incubated for 30 min at 55°C cDNA;mRNA hybrids were purified with QIAGEN Mini Elute PCR Purification Kit. All subsequent steps are according to the CEL-seq2 method as follows: Second strand DNA is synthesized by nick translation method using RNase H, DNA polymerase I, and DNA ligase mix, by which only the cDNAs derived from the photo-irradiated region are filled-in to the 5' end of T7 promoter, thereby allowing the cDNAs to be linearly amplified as RNAs (aRNAs) by IVT reaction. The libraries are further reverse-transcribed, followed by paired-end sequencing on the Illumina platform.