3 RACE TAF1 ex30-polyA - PFC
3 RACE was performed using published procedure (PMID: 17406530). Briefly, cDNA synthesis was primed with a modified oligo(dT) primer (QT) which selectively target polyadenylated transcripts and adds two specific sequences, QO and QI, for subsequencent amplification. PCR amplification was carried with PrimeSTAR GXL DNA Polymerase (Takara Bio) using a gene-specific forward primer located at the beginning of TAF1 exon 30 in combination with QO reverse primer. A subsequent nested reaction increased specificity by using an inner forward primer located at the end of TAF1 exon 30 in combination with QI. Amplified products were purified with AMPure XP Beads (Beckman Coulter) and further processes for Nanopore long-read sequencing using the kit SQK-NBD114.24 or SLK114 combined with the barcoding kit EXP-PBC-096 according to manufacturers instructions. Libraries run on MinION device, mounting MinION or Flongle flow cells for 24-72 h.
SUB15539743
17.17