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Bacteriophage Tail Fibers as a Basis for Structured Assemblies by Timothy Harrah Harrah, Paul Hyman

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Appendices

A. Tail fiber purification

Crude lysate

An overnight culture of Bb on 2×YT (with 0.2% glucose) was used to make a 1% inoculum in a 20 L fermentor (GRASP center, NEMC, Boston). The culture was aerated at 37°C to a cell density of 5×108/mL and infected with either T4 9am tam (for LTF) or T4 9am 37S∆1 tam (for S∆1 LTF) at an MOI of 5. The culture was incubated for 2 hours, chilled immediately on ice to prevent premature cell lysis and centrifuged at 6,500 g (Sorvall, rotor GSA) for 10 minutes at 4°C. The cell pellet was gently resuspended in XB at 100× concentration and lysed with CHCl3 at room temperature for 30 minutes with gentle shaking. This lysate was centrifuged at 11,000 g for 10 minutes (Beckman, JA 20 rotor).

This crude cell ...

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