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Total RNA was extracted and assessed for quality (RIN > 7.0). Sequencing libraries were prepared using the Illumina TruSeq Stranded mRNA Library Prep Kit. Briefly, poly(A) mRNA was purified from total RNA using oligo(dT) beads, fragmented, and reverse-transcribed into first-strand cDNA. Second-strand synthesis incorporated dUTP to quench amplification of the second strand, preserving strand specificity. After end repair, A-tailing, and adapter ligation, libraries were enriched by PCR, purified, and validated on a Bioanalyzer.Indexed libraries were pooled and sequenced on the Illumina NovaSeq 6000 platform with paired-end 150 bp (PE150) reads. |
RNA-Seq |
TRANSCRIPTOMIC SINGLE CELL |
RANDOM |
PAIRED
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