GSM7990931: Definitive Endoderm cells, saracatinib 1; Homo sapiens; RNA-Seq
Initially, messenger RNA (mRNA) was isolated from 1 μg of total RNA using oligo (dT) magnetic beads, followed by a fragmentation process using divalent cations at elevated temperatures in the ABclonal First Strand Synthesis Reaction Buffer. First-strand cDNAs were synthesized using random hexamer primers and Reverse Transcriptase (RNase H), with the mRNA fragments serving as templates. This was followed by the generation of second-strand cDNAs using DNA polymerase I, RNaseH, buffer, and deoxyribonucleotide triphosphates (dNTPs). The resulting double-stranded cDNA fragments were then ligated with adapters to prepare the paired-end library.