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PEG precipitation

SH Sarah L. Hulin-Curtis
JD James A. Davies
DN Davor Nestić
EB Emily A. Bates
AB Alexander T. Baker
TC Tabitha G. Cunliffe
DM Dragomira Majhen
JC John D. Chester
AP Alan L. Parker
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The PEG precipitate was centrifuged at 10,000 rpm for 15 min at 4 °C. The supernatant was discarded and re-centrifuged briefly to remove residual supernatant. The phage pellet was resuspended in 1 mL of TBS and centrifuged in a clean tube at 13,000 rpm, for 5 min at 4 °C. The supernatant was transferred to a clean tube, precipitated with 1/6th volume (~166 µL) of PEG/NaCl, incubated on ice for 30 min followed by centrifugation at 13,000 rpm for 10 min at 4 °C. The supernatant was discarded and centrifuged briefly to remove residual supernatant. The pellet was resuspended in 200 µL of TBS, 0.025 NaN3 and centrifuged for 1 min to pellet residual insoluble material. The supernatant was transferred to a clean tube as the amplified eluate. The amplified eluate was titred as described above using 108–1011 dilutions of amplified phage eluate.

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