Author: Feng, Mingqian; Bian, Hejiao; Wu, Xiaolin; Fu, Tianyun; Fu, Ying; Hong, Jessica; Fleming, Bryan D; Flajnik, Martin F; Ho, Mitchell
Title: Construction and next-generation sequencing analysis of a large phage-displayed V(NAR) single-domain antibody library from six naïve nurse sharks Document date: 2018_11_7
ID: wc6k06sm_31
Snippet: Library TG1 bacterial stock was inoculated into 2.5 L of 2YT media containing 2% glucose, 100 μg/ml ampicillin, and cultured at 37 • C with shaking (250 rpm). When the cells reached mid-log phase (OD 600 between 0.4-0.8), super-infection was performed by adding helper phage M13KO7 at 5 × 10 9 pfu/ml. After 1 h of continued growth, the TG1 cells were pelleted and resuspended in 2.5 L of 2YT media containing 100 μg/ml ampicillin and 50 μg/ml .....
Document: Library TG1 bacterial stock was inoculated into 2.5 L of 2YT media containing 2% glucose, 100 μg/ml ampicillin, and cultured at 37 • C with shaking (250 rpm). When the cells reached mid-log phase (OD 600 between 0.4-0.8), super-infection was performed by adding helper phage M13KO7 at 5 × 10 9 pfu/ml. After 1 h of continued growth, the TG1 cells were pelleted and resuspended in 2.5 L of 2YT media containing 100 μg/ml ampicillin and 50 μg/ml kanamycin, and incubated at 25 • C overnight. After the cells were centrifuged and filtered with a 0.22 μm membrane, the supernatant was stored at 4 • C for panning.
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