Shark PBL RNA Extraction and cDNA Library

Total RNA was extracted from the shark’s PBLs and used as templates to synthesizing first-strand cDNA using oligo(dT) primers. The library encoding sequences were amplified by PCR from cDNA, the framework specific primers Bam VF1 CGCGGCCCAGCCGGCCATGGCCGCCSMACGGSTTGAACAAACACC and Bam VF2 CGCGGCCCAGCCGGCCATGGCCGCCGCACGGGTTGAACAAACACCG. DNA fragments were cleaved with restriction enzymes Nco I and Not I (NEB) for use in subsequent experiments. An anti-OGT phage display library of about 10⁸ independent transformants was obtained following the detailed protocol as Ubah et al. (27 (link)).