GSM8758984: Neuroblastoma xenograft derived from SK-N-BE(2) cells treated by RA+NEN diet (Bio-replicate 2); Homo sapiens; RNA-Seq
Tumors were dissociated using the Tumor Dissociation Kit (Miltenyi; #130-095-929) in combination with the gentleMACS™ Octo Dissociator (Miltenyi; #130-096-427). The resulting single-cell suspensions were prepared for library construction using the PIPseq™ V T10 3' Single Cell RNA Kit. The PIPseq™ T20 3ʹ Single Cell RNA Kit v4.0PLUS protocol begins with isolating single cells and lysing them in the provided buffer. Poly(A)+ RNA is captured using oligo(dT) primers with unique molecular identifiers (UMIs), followed by reverse transcription and template switching to generate full-length cDNA. The cDNA is amplified via PCR, purified using magnetic beads, and fragmented with a transposase for library preparation. Unique dual indices (UDIs) are added during a final limited-cycle PCR to enable multiplexing. Libraries are then purified, quality-checked using a bioanalyzer or TapeStation, and quantified by Qubit or qPCR before sequencing on an Illumina platform.