title: Site saturation mutation of TrkA binders 7154&4240
design: Yeast plasmids were extracted (TIANprep Yeast Plasmid DNA Kit) and eluted in 70 l distilled water, serving as templates for deep sequencing. Deep sequencing libraries with library-specific barcodes were prepared by PCR amplification for 2230 cycles in a 50 l KOD One reaction system. The PCR products were separated on a 1.5% agarose gel and extracted (ComWin Biotech Gel Extraction Kit). Purified DNA were mixed and sent for deep sequencing (Novogene).