title: Single-cell and spatial transcriptomic analyses revealing tumor microenvironment remodeling after neoadjuvant chemoimmunotherapy in non-small cell lung cancer
design: Use universal primers at both ends of the first strand cDNA to perform PCR amplification, synthesize the second strand cDNA, increase the cDNA content, and then purify the product, which can be used for subsequent T cell enrichment libraries and 5' gene expression libraries 's build. Design highly specific primers in the conserved regions of TCR or Ig, and enrich barcoded full-length V(D)J fragments from cDNA by nested PCR.Before constructing the library, the enriched V(D)J fragments were digested and screened to generate DNA fragments of different lengths. The V(D)J fragment has R1 (read 1 primer sequence) added by GEM-RT reaction, P5 added when the target gene is enriched, and P7 added by end repair, A tail, adapter ligation and PCR during library construction ( sample indicator) and R2 (read 2 primer sequence). The final constructed library product contains P5 and P7 sites and can be used for Illumina bridge amplification.
identifiers.namespace: SUB15307802
identifiers.id: J22100183