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Blocking solution: 5 % (w/v) skimmed milk in PBS. 7. Pure streptavidin dissolved in PBS at 2 mg/ml concentration. 8. 05 % (v/v) PBS-T. 9. 01 % (v/v) PBS-T. 10. HRP-conjugated Pittsburgh, PA). anti-M13 antibody (GE Healthcare, 11. 01 % (v/v) PBS-T. 12. TMB substrates and 5 N H2SO4. 2 DNA Sequence Analysis 1. 10× PCR buffer, MgCl2, dNTP, and Taq polymerase: Expand High Fidelity PCR system (Roche, Indianapolis, IN, USA). 2. PCR primers: 5′-CGGCGCAACTATCGGTATCAAGCTG-3′. 5′-CATGTACCGTAACACTGAGTTTCGTC-3′.

Such libraries are created by inserting random DNA sequences at an appropriate location within a structural gene of a phage particle. Peptides with affinity for a target protein may then be selected through cycles of selection and amplification of phages. The most commonly used vectors for phage display are Ff filamentous Escherichia coli bacteriophage, which infects E. coli using the F-pilus as the primary receptor. Without undergoing lysis, infected bacteria produce and secrete the phage particles, which are about 65 Å in diameter and nearly 1 μm in length.

EcoRI, BglII, and 10× NEBuffer EcoRI (New England Biolabs). 3. SeaKem GTG Agarose and NuSieve GTG Agarose (Fisher Scientific). 4. 0. Store at room temperature. 5. Gel casting trays for agarose gel electrophoresis of approximate size 20 cm × 30 cm and 10 cm × 10 cm, with combs that contain one broad well for application of large samples flanked by small wells for molecular weight marker DNA. 6. 25 % (w/v) bromophenol blue, 30 % (v/v) glycerol. Store at room temperature. 7. Ethidium bromide: 10 mg/mL stock solution.

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