GSM8283786: Cytoplasmic region of mock-infected A549 cells; Homo sapiens; OTHER
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℃. cDNA;mRNA hybrids were purified with a Qiagen MinElute PCR Purification kit. Library was constructed according to the CEL-seq2 method as follows: polyA RNAs were converted in situ to cDNAs using a polyT primer with a 5'-tail containing T7 promoter, 5' Illumina sequence adaptor, UMI and cell barcode sequences. The cDNAs were extracted and second strand DNAs were synthesized by nick translation method. The cDNAs were linearly amplified as RNAs (aRNAs) by IVT reaction, followed by reverse transcription using random primer with a 5'-tail containing 3' Illumina sequence adaptor. Spatial transcriptome analysis using PIC (photo isolation chemistry).  PIC is a bulk RNA-seq technique to prepare cDNA libraries from specific regions (not cells) in tissue sections.