GSM5870206: Pat 4 Visit 1 SF left Treg; Homo sapiens; RNA-Seq
Total RNA was isolated using the RNeasy Mini Kit (Qiagen) for cell fractions ≥0.2 million cells and the RNeasy Micro Kit (Qiagen) for fractions ≤0.2 million cells, following the manufacturer's instructions. cDNA was synthesized using the SMARTer RACE cDNA Amplification kit (Clontech). Amplification of the TCRβ VDJ region was performed using previously described primers and amplification protocols(Zhou et al., 2006). PCR product fragment size was analyzed using the QIAxcel Advanced System (Qiagen). End repair and barcode adapter ligation were performed with the NGSgo®-LibrX and NGSgo®-IndX (GenDx) according to the manufacturer's instructions. Cleanup of the samples was performed after each step using HighPrep PCR beads and following the manufacturer's instructions (GC Biotech).