Figure 2.
A mock selection cycle using stapled mRNA display to measure enrichment of E2 binding peptide 10. P = puromycin. mRNAs corresponding to peptide 10 and the library mRNAs were each ligated to a puromycin DNA sequence. These puromycin-mRNAs were then added to in vitro translation reactions, leading to fusion of the nascent linear peptides onto their encoding mRNAs. Each mRNA-peptide fusion was then cyclized with DBX, followed by reverse transcription to generate the cDNA. The mRNA-peptide fusions were then captured onto Avi-tagged HPV16 E2.44 The non-binding mRNA-peptide fusions were washed away, and binders were then eluted with heat and amplified by PCR.